Download C7. Time schedule of the research plan solution

RP identification code
MSM0021620812
Research plan proposal
Provider‘s code
MSM
RP identification code 0021620812
Research plan title
Chronic diseases originated from inappropriate reactivity of immune
system, their pathogenesis and possibilities of their early diagnosis
and treatment.
Applicant
Charles University, Prague
Administrator
2nd Medical School
Investigator
Prof. Jirina Bartunkova, MD, DrSc.
A
A1.
Principal information on the applicant
Organisational scheme of the applicant indicating the number of employees
Charles University in Prague is a public institution which has been in operation since its foundation on
the 7th of April 1348.
It comprises 17 faculties, 4 institutes of higher learning, 6 other workplaces and 5 special-purpose
establishments.
(The executive body of the university is the Rectorate. Details are specified by the Charles University
Statute and the organizational structure by Annex No. 2 - Code of Organisation
Faculties:
Catholic Theological Faculty
Protestant Theological Faculty
Hussite Theological Faculty
Faculty of Law
1st Faculty of Medicine
2nd Faculty of Medicine
3rd Faculty of Medicine
Faculty of Medicine in Pilsen
Faculty of Medicine in Hradec Králové
Faculty of Pharmacy in Hradec Králové
Faculty of Arts
Faculty of Natural Sciences
Faculty of Mathematics and Physics
Faculty of Education
Faculty of Social Science
Faculty of Physical Education and Sport
Faculty of Humanities
Number of staff and employees
63
64
102
258
1660
556
628
472
450
231
740
785
770
503
239
309
91
University Institutes:
Institute of the History of CU and CU Archives (Rectorate)
Centre for Theoretical Studies (Rectorate)
Centre for Economic Research
and Graduate
59
Other University Workplaces:
Computer Centre (Rectorate)
European Information Centre (Rectorate)
Institute of Language and Specialist Training
Central Library (Rectorate)
Education
178
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MSM0021620812
Environmental Centre (Rectorate)
Agency of the Council of Higher Education Institutions (Rectorate)
Special-Purpose Establishments:
Halls of Residence and Refectories
Archbishop’s Seminary
The Karolinum Press (Rectorate)
Management of Buildings and Facilities
Sports Centre (Management of Buildings and Facilities)
Rectorate
818
12
113
303
Overall number of Charles University Staff and Employees In Prague
2
9404
RP identification code
MSM0021620812
A2.
Bodies pertaining to the applicant and their personnel
According to the Law No. 111/1998 Coll. on Institutions of Higher Learning and Statutes of Charles
University in Prague the organs of Charles University in Prague are:
1. Independent academic bodies
a) Academic Senate (for member list see the Annex A2-1)
b) Chancellor - prof. Ing. Ivan Wilhelm, CSc.
c) Scientific Council (for member list see the Annex A2-2)
The Disciplinary Commission of Charles University has not been established (§13, law. No. 111/1998
Coll., on Institutes of Higher Learning)
2. Other bodies
a) Board of Administration (for member list see the Annex A2-3)
b) Bursar - Ing. Josef Kubíček
A2-1
Mgr. Václav Balek
Doc. RNDr. Jiří Banýr, CSc.
PhDr. Petr Bednařík, Ph.D.
John Bekkenes
RNDr. Štěpán Bojar
Peter Brezina
Jana Buchtová
Doc. MUDr. Zuzana Červinková, CSc.
Prof. PhDr. Marie Dohalská, DrSc.
Doc. PharmDr. Martin Doležal, Ph.D.
Tomáš Drbohlav
MUDr. František Duška
Dr. Karel Fajfrlík
Daniel Feranc
Jan Foniok
RNDr. Daniel Frynta, Ph.D.
Pavel Gonda
Prof. MUDr. Pavel Gregor, DrSc.
Doc. MUDr. Petr Hach, CSc.
Prof. RNDr. Jan Hála, DrSc.
Prof. RNDr. Václav Hampl, DrSc.
J. Hodan
Martina Hrubešová
Petr Chovanec
MUDr. Zuzana Jarkovská
Lukáš Kielberger
PhDr. Jiří Kirchner
Kateřina Kočvarová
Ondřej Kobza
Ondřej Konvalina
Martin Kopta
Mgr. Jan Kranát, Ph.D.
JUDr. Michael Kučera
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Mgr. Miroslav Kudláček
Prof. PhDr. Jan Kuklík, CSc.
Doc. RNDr. Ladislav Lešetický, CSc.
Doc. PharmDr. Miloš Macháček, CSc.
Doc. MUDr. Bohuslav Matouš, CSc.
Ing. Petr Mikeš
Mgr. Jakub Mrázek
Doc. PhDr. Viléma Novotná
Jan Pavlík
Mgr. Jiří Pavlík
Michal Pelíšek
Mgr. Zdeněk Pressl
PhDr. Libor Prudký
PhDr. Irena Reifová, Ph.D.
Doc. RNDr. Josef Reischig, CSc.
Doc. PhDr. Ing. Jan Royt, CSc.
Oldřich Řeháček
Jan Říčař
Ján Říha
JUDr. Mgr. Josef Salač, Dr.
Th. D. Petr Sláma
Tomáš Slavík
Tomáš Soukup
JUDr. Ing. Josef Staša, CSc.
PhDr. Josef Stracený, CSc.
Prof. MUDr. Pravoslav Stránský, CSc.
Doc. MUDr. Richard Škába, CSc.
Adam Šůra
Mgr. Daniela Tinková, Ph.D.
Jan Tuček
Zdeněk Turek
Doc. MUDr. Michael Urban, CSc.
Pavel Vychytil
Ing. František Zahálka, Ph.D.
Doc. RNDr. Miloš Zahradník, CSc.
PhDr. Filip Žikeš
A2-2
Prof. Ing. Ivan Wilhelm, CSc.
Prof. Pavel Ambros, Th.D.
Prof. MUDr. Jan Betka, DrSc.
Prof.PhDr. Petr Blahuš, DrSc.
Prof. MUDr. Jan Bubeník, DrSc.
Prof.MUDr.Josef Fusek,DrSc.
Prof.PhDr.RNDr.Helena Haškovcová, CSc.
Doc. PhDr. Vilém Herold, DrSc.
Doc. RNDr.Helena Illnerová, DrSc.
Prof.MUDr. Richard Jelínek, DrSc.
Ing. Karel Jungwirth, DrSc.
Prof.RNDr. Rolf Karlíček, DrSc.
Prof. MUDr. Jan Kilian, DrSc.
Prof.MUDr.Pavel Klener, DrSc.
Prof. PhDr. Jiří Kraus, DrSc.
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Prof. ThDr. Zdeněk Kučera
Ing. Miroslav Kuchař, DrSc.
Prof. PhDr. Jan Kuklík, CSc.
Prof. PhDr. Jiří Kuthan, DrSc.
Prof. MUDr. Jan Libiger, CSc.
Prof. Ing. Miroslav Ludwig, CSc.
Prof. Ing.Josef Macháček, DrSc.
Prof.JUDr.Karel Malý, DrSc.
Prof. PhDr. Jiří Mareš, CSc.
Prof.MUDr. Jaroslav Masopust, DrSc.
Prof. Ing. Lubomír Mlčoch, CSc.
Prof. JUDr. Jan Musil, CSc.
Prof.MUDr.Bohuslav Ošťádal, DrSc.
Prof.RNDr. Václav Pačes, DrSc.
Prof. RNDr. Zdeněk Pertold, CSc.
Doc.RNDr.Petr Pikálek, CS.
Prof. RNDr.Jaroslav Pokorný, CSc.
Prof.ThDr.Petr Pokorný, DrSc.
Doc. RNDr. Josef Reischig, CSc.
Prof. Ing. Josef Rosenberg, DrSc.
Doc.RNDr. Vladimír Semecký, CSc.
Doc. PhDr. Jan Slavík, CSc.
Doc. MUDr.Jan Starý, DrSc.
Prof.RNDr.Václav Suchý,DrSc.
Doc. RNDr.Jaroslava Svobodová, CSc.
Prof.MUDr.Josef Syka, DrSc.
Prof. PhDr. František Šmahel, DrSc.
Prof.RNDr.Josef Štěpán, DrSc.
Prof. Ing. Karel Štulík, DrSc.
Prof. JUDr. Jiří Švestka, DrSc.
Prof.ThDr. Ing. Jakub Trojan
Doc. Ing. Zdeněk Tůma, CSc.
Prof. PhDr. Miroslav Verner, DrSc.
Prof. MUDr. Zbyněk Vobořil, DrSc.
Prof. MUDr. Petr Widimský, DrSc.
Prof. MUDr. Tomáš Zima, DrSc.
A2-3
JUDr. Antonín Mokrý
MUDr. Pavel Bém
Ing. Oldřich Dědek, CSc.
JUDr. Ing. Miloslav Fiedler
Karel Jan Jeníček
JUDr. Jan Kollert
PhDr. Miroslav Kostka
Ing. Vratislav Kulhánek
JUDr. Jiří Srstka
Doc. Dr. Milan Stloukal, DrSc.
PhDr. Miloslav Vlk
Doc. Ing. Jiří Volf, CSc.
JUDr. Jan Wagner
PhDr. Lubomír Zaorálek
Prof. Ing. Petr Zuna, DrSc.
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RP identification code
MSM0021620812
A3.
Overview or profile of all activities pursued by the applicant
The mission and activities of the university (applicant) are defined by the article 2 of the Statute giving
their basic outline.
The applicant (university) has been affecting educational activities in accredited study
programmes (bachelor’s, master’s, doctoral), as well as in the lifelong education programmes.
Furthermore, it affects scientific and other research, development and other creative
activities. All these basic activities are carried out on an international scale and on various levels of
cooperation.
The university provides various kinds of information services as well as library services; at the
same time it pursues publication and consultation activities.
Within the framework of its expertise the University carries out specialist and consultation
activities.
Within the framework of its internal administration the university manages organisational,
coordination, consultative, research and supervisory activities in various fields of study including
science, economics, personnel, legislative and external relations including foreign contacts.
Further, activities are carried out on the university premises according to the Trade Law as listed
below:
Free Trade
Automatic processing and data transmission,
Biology tests on laboratory animals,
Technical consultations in the field of Physics,
Economic and organisational consultations,
Expert activities based in analyses of processing, information, commentaries, studies and of the
living environment projects,
Copy service,
Commodity purchase with the aim of selling, and sales
Radon measurements,
Book Publishing,
Procurement of activities in the administration of non-residential real estate,
Consulting for Industry and Commerce,
Courses and Seminars in the field of Humanities,
Courses and Seminars in the field of Social Sciences,
Education and Instruction ventures,
Exhibition, fairs, displays, shows and similar ventures,
Provision of services for personal hygiene and services related to image and beauty treatment,
Software provision,
Rental of non-residential premises including other than basic services assuring proper operation of
the non-residential premises,
Lease of sports facilities,
Running galleries (except the activities excluded by the law No. 455/91 coll., in the wording of
later directives),
Translation and Interpreting services,
Psychological consultations and diagnostics,
Advertising and commercial campaigns,
Accommodation services,
Magazines and Journals Publishing,
Chemicals and chemical formulations production, – chemical substances and chemical
preparations for laboratory and manufacturing purposes,
Production of optical components,
Production, copying, multiplication and recording of audio, audio and video recordings,
Production of printing templates
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Foreign languages teaching,
Czech language teaching,
Research and development in the field of natural science and technical sciences or social sciences
– analysis, application and realisation of algebraic algorithms,
Processing of expertise and opinion in the field of art and social sciences,
Business mediation,
Advertisement mediation.
Regulated trade and craft
Geological work,
Hostelry activities,
Massage, reconditioning and regeneration services,
Printing,
Physical education services,
Driving School,
Travel Agency,
Provision of physical education, regeneration and reconditioning facilities,
Joinery.
Licensed trade
Haulage,
Road passenger transport,
Telecommunication services.
A4.
Share of research and development in the total activities of the applicant
Indicator
2001
2002
Number of employees of the applicant engaged in research and
1146
1188
development
Number of FTE adjusted work loads implemented in research and
803,9
797,4
development
Share of expenditures on research and development in the overall
22,3
19,8
expenditures of the applicant in per cent*)
*) Or some other indicator, which financially stipulates the share of research and development in the
overall activities of the applicant, may be used
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B
B1.
Information on research and development activities of the
applicant/administrator
Specification of the principal research and development activity of the
applicant/administrator
Scientific work represents one of the central points of the Faculty activities. It contributes to
its high standards and its status on an international level. A few internationally respected
groups work at the 2nd Medical Faculty - Charles University, both in the field of basic and
applied clinical research. The quality is also enhanced by our success in getting grants
financed by various grant agencies and also by high-quality publications issued at the Faculty.
In recent years the quality of publications has improved and primary publications, with high
impact factor issued in foreign periodicals, start to be quite common. Successful
implementation of grant projects also proves high standard of our scientific work. All the
projects were completed without any comments. Another important qualitative change, in the
field of faculty research work, is the integration of laboratory and clinical research activities
which ensue from a number of favorable factors. Thanks to scientific activities, financed
from grants, research groups (laboratories), which are very efficient, are constituted in
departments. Another positive factor which contributes to stimulation of qualified research
work, is the postgraduate program of further scientific education. The postgraduate study
program begins to prove its effectivity, the number of successfully defended PhD thesis is
gradually rising. The Faculty is a part of a common postgraduate education program of all
medical faculties in Prague, the Faculty of Science, the Czech Academy of Sciences (AS CR)
and departmental research workplaces. Project oriented financing of research plans also plays
an important role. At present, five research plans are carried out at the Faculty and they are all
evaluated approvingly. At the same time they significantly support team work. There are three
research centers existing at the Faculty bringing the amount of 25 million Czech crowns for
faculty research activities. The 2nd Medical Faculty belongs, in this field, to the most
successful ones and the Faculty maintains its good position in Czech medical science.
Research centers play an essential role in education of young scientific generation.
B2.
Contribution of the applicant/administrator to the development of knowledge in
the disciplines referred to in B1, on national and international scale
The scope of scientific activities of the 2nd Medical Faculty and Teaching Hospital in Motol
is wide and covers many issues both from the sphere of clinical research and basic
biomedical research. Centrally run and generally accessible national and international
publication databases keep the documentation, therefore we do not consider it necessary to
specify this point in further details. We focus on outstanding results in the particular fields
which the Faculty intends to involve in further research plans.
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B3.
Major research and development results applied by the applicant/administrator
in the disciplines referred to in B1 within the last five years (overall characteristic)
As far as immunology is concerned, it has a long tradition at the 2nd Medical Faculty, it has
already existed since the 1970´s. The discipline has quickly developed, both in clinical and
research fields, especially after establishing the Department of Immunology in 1995 - a
common department of the 2nd MF and Motol TH. In cooperation with other workplaces
several priorities have been achieved on national and international levels. Among these are
for example, cord blood transplantation in a patient with a rare immunodeficiency syndrome
and the first prenatal diagnostics. Results achieved in research sphere were published in
internationally reviewed magazines, e.g. in a review summing up many years´ experimental
and clinical work in ANCA-positive vasculitides (2) and the first publication in the Czech
Republic, dealing with methodology of cancer vaccine preparations on the basis of dendritic
cells (3). According to results both in treatment preventive care and in research field, the
American Federation of Clinical Immunology Societies (FOCIS) acknowledged the 2nd and
1st Medical Faculties as one of the international centers of this association. This centre
represents the basis for coordination of immunologically directed research, which is the core
of the presented research plan for 2005 – 2010, and which coordinates important research
immunological groups at medical faculties of Charles University into one functional unit.
1. Starý J, Bartůňková J, Kobylka P, Vávra V, Hrušák O, Calda P, Král V, Švorc K.
Successful HLA-identical sibling cord blood transplantation in a 6-year-old boy with
leukocyte adhesion deficiency syndrome. Bone Marrow Transplantation 1996; 18 (1):
249-252.
2. Bartůňková J., Tesař V., Šedivá A.: Diagnostic and pathogenetic role of antineutrophil
cytoplasmic antibodies. Clinical Immunology 10,2003: 73-82.
3. Pospíšilová D., Borovičková J., Poloučková A., Špíšek R., Šedivá A., Hrušák O.,
Starý J., Bartůňková J.: Generation of functional dendritic cells for potential use in the
treatment of acute lymphoblastic leukemia. Cancer Immunology Immunotherapy
51,2002,2:72-78.
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B4.
International cooperation of the applicant/administrator in research and
development
B4.a. Participation of the applicant/administrator in international research and
development cooperation implemented on the basis of international contracts
concluded by the CR with foreign entities
The 2nd Medical School UK cooperates with foreign partners in the following projects based
on interstate contracts: Erasmus/Sokrates, Marie Curie, Smartie.
B4.b. Collective membership of the applicant/administrator in non-governmental
international research and development organisations
Collective membership is carried out in professional associations.
Czech Society of Allergology and Clinical Immunology is integrated in international EAACI
and WAO programs. Czech Immunological Society is a member of IUIS.
Czech immunology of reproduction is a part of European and international organization (AlpAdriatic Society for Reproductive Immunology, European Reproductive and Developmental
Immunology, International Society for Reproductive Immunology).
B4.c. Individual membership of employees of the applicant in non-governmental
international research and development organisations
Reported in all research workers individually.
B4.d. Contracts or joint projects of the applicant/administrator with foreign
organisations engaged in research and development
Allostem (The Development of Immunotherapeutic Strategies to Treat Haematological and
Neoplastic Diseases on the Basis of Optimised Allogeneic Stem Cell Transplantation), the 6th
RP EU , contract number 503319 – CR coordinator: J. Bartůňková
EURO-PID (QLTR-2001-02742), research of primary immunodeficiencies, in the framework
of the 5th RP EU, A. Šedivá: coordinator
Erasmus – cooperation with Karolinska Institute in Stockholm, Sweden - Dendritic Cells
Immunotherapy project. Exchange programmes for members of pedagogical staff and
postgraduate students, common conferences (coordinators: J. Bartůňková on behalf of the 2nd
MF CU, G. Masucci on behalf of Karolinska Institute).
B4.e. Other forms of international cooperation
Cooperation with Hopital Necker Enfants Malades in Paris: diagnostics and treatment of
primary immunodeficiencies, common postgraduate programme (J. Bartůňková, A. Šedivá).
Participation in an international project „EUVAS“ – a European Vasculites Group,
international multicentric studies (J.Bartůňková).
Participation in postgraduate education programme and immunotherapy in the framework of
La ligue contre le cancer with a French Institute de Biologie, Nantes (R. Špíšek).
Participation in the MiniMini project: Research Section, Pediatric Hematology/Oncology,
Sheba Medical Center, Tel Hashomer, Israel; Institute of Immunology, Zagreb, Croatia (O.
Hrušák).
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Cooperation in BFM/AEIOP 2000 report: Children´s Cancer Research Institute, St Anna
Kinderspital, Vienna, Austria; Pädiatrische Hämatologie/Onkologie, Universität Klinikum
Münster; detection of minimal residual illness in acute myeloid leukemia in the framework of
„MPWG“ study – Münster, Prague, Vienna, Göttingen (O.Hrušák).
Cooperation with Prof. J.J.M. van Dongen, Rotterdam, in immunodeficiency
and leukemia diagnostics.
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B5.
Research and development budget of the applicant/ administrator in the previous
five years in thousands of CZK
Institutional
Targeted
Specification of
Year
support from
support from Other resources
other resources
the State Budget the State budget
1998
33865
1948
638
EU grants
1999
44978
8507
1889
-“2000
53061
13140
2687
-“2001
53870
13223
2486
-“2002
67671
13197
7138
-“-
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C
C1.
plan
Research plan description
Specification of the research activity subject implemented within the research
Disorders of the immune system represent a serious problem of public health. An insufficient
or an exaggerated function of the immune system causes allergic, autoimmune, primary and
secondary immunodeficiencies and leads to the development of tumor diseases. Recent
estimates claim that as many as 40% of the population suffers from a disease that results from
an inappropriate reactivity of the immune system.
The proposed research plan aims at studying such common basic mechanisms that lead to
development of the above mentioned disorders and concentrates on possibilities of their early
diagnosis and therapeutic modulation. Specifically, we plan to study the following:
 the influence of specific genetic factors associated with the disorders of the immune
mechanisms
 functional imunopathologic aspects leading to the damage of the organism
 laboratory markers enabling early diagnosis of immunopathologic states
 use of immunotherapy and biological therapy in the treatment of immune disorders
The project is conceived as a close cooperation between departments of immunology of the
medical faculties at the Charles University, with additional participation of clinical and
theoretical departments of other institutions according to particular subprojects. Both training
and education of PhD students and postdocs shall represent an integral part of this project.
C2.
Present level of knowledge and research activity, which is the subject of the
research plan, from both international and national standpoints
Immunology, initially considered as a purely experimental and theoretical field, has reached
an incredible boom and importance in the past decade and it has become a key clinical
speciality. It is not exaggerated to claim that at present all medical specialities use
immunological knowledge either in the process of diagnosis or for the treatment. Disorders of
the immune system can manifest as an impairment of all organ systems. Insufficient or
exaggerated functions of the immune system may cause allergic, autoimmune, primary and
secondary immunodeficiencies and lead to development of a number of tumor diseases.
Recent estimates claim that as many as 40% of the population suffer from a disease that
results from the inappropriate reactivity of the immune system. Thus, such disorders represent
a major problem of public health. A group of such disorders is now frequently designated as
Immune-mediated inflammatory disorders (IMID). In the past, research groups in
immunology concentrated on specific, highly specialized, models of immunopathology. It is
not a surprise, because immunity disorders are represented by such different diseases as
allergic rhinitis, bronchial asthma, atopic dermatitis, type I. diabetes, Hashimoto´s thyroiditis,
systemic lupus erythematosus, rheumatoid arthritis, vasculitis, sclerosis multiplex, immunemediated disorders of reproduction, insufficient immune response leading to tumor diseases,
immunopathological reaction during sepsis and polytrauma, immunodeficiencies in chronic
infections, primary immunodeficiencies, anaphylaxis, and many more. However, in the course
of the time, it became obvious that the disorders of the immune system shared many common
aspects and that results obtained in one experimental model could be transferred and exploited
in other diseases. Primary immunodeficiencies could serve as a model example. In this group
a unique well-defined genetic lesion can lead, in one patient, to symptoms of
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immunodeficiency, autoimmunity, allergy or even tumor disease. Similarly, HIV infection has
an analogous spectrum of diverse symptoms. A majority of immune disorders is polygenic
and the experimental models help to identify the groups of genes associated with certain
immunopathology. At the present time, there is a worldwide tendency to concentrate
individual research groups into complex multi-task teams. Then, every research team
investigates a concrete, well-defined problem, and a coordination of their results accelerates
an application of new findings in clinical practice. The FOCIS Center (www.focisnet.org),
USA, brings together research teams from USA, Canada and Western Europe, focusing on
projects in clinical immunology. Our immunologic group has recently become a member of
this international consortium. Social relevance of immunopathologic disorders is well
documented by the fact that the most prestigious research institute in the USA-NIH- spends
several billions of American dollars per year on projects concerning diseases associated with
immune disorders. This example underlines the importance of targeted support of
immunology in the Czech Republic, especially in the field of applied research, that ensures
applications of new important experimental findings in clinical practice.
Immunology in the Czech Republic has a long tradition. The basic research has concentrated
at institutions of the Czech Academy of Sciences, and the applied projects have been
investigated at the medical faculties. Currently, there is a tendency to closely link these two
major fields of research and the medical faculties are the best candidates for the establishment
of such centers. The team that presents this proposal is composed of members of
immunological departments of the medical faculties at the Charles University. They all have
longstanding experience in either basic or clinical immunology. The research team at the
Department of Immunology of the 2nd Medical Faculty at the Charles University, the principle
investigator of this project, has achieved a number of priority results in both national and
international context (see section b3). The prestige of the particular immunological
departments is well documented by publications and research activities outlined in individual
investigators. According to the complexity of immune mediated disorders, individual
departments have gradually focused on particular fields of immunopathologies. That is why
this proposal is structured into distinct subspecialties of clinical immunology and each section
will be coordinated by the center that has achieved the most important results. Similarly to the
tendency in the USA and Western Europe, research activity of all teams will be concentrated
in a functional entity. Thus, it will be possible to apply and use the results presented by each
group for the activity of remaining subjects, and respect the common ethiopathogenetic
mechanisms of IMID. As an advantage of such coordination of the research effort of all
participating groups within one project, costs of the research efforts shall be reduced, as this
organization structure permits to check and avoid any duplicities in financing and helps with
an investments coordination.
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C3.
Relationship between of the research plan and the research specialisation of the
applicant/administrator and its connection to the long-term perspective of
research and development of the applicant/administrator
An extremely efficient research group has developed at the Department of Immunology at the
2nd Medical Faculty, Charles University, since 1995. This group has, in cooperation with
others, achieved a number of priority findings in both national and international context (see
section b). A similar situation exists at other medical faculties. The administration and
management in all the medical faculties involved in this proposal support the research in the
fields of immunology, given the benefits of the immunology issues for the society, as a whole.
C4.
Current share of the applicant/administrator in the solution of the subject of the
research plan on both national and international scales
Research teams involved in this proposal are respected in the field of immunopathology at the
national and international level. At the national level, this can be documented by the
membership of investigators in the committees of various relevant societies (Czech Society
for Alergology and Clinical Immunology, Czech Immunology Society, Society of
Rheumathology), in the review boards of grant agencies (GACR, IGA, GAUK), and in
editorial boards of Czech scientific journals (Alergologie, Revmatologie). Investigators are
the authors of the immunology textbooks and they participate at the development of
therapeutic guidelines in immunology. At the international level, the most important is a
continuous publication activity in respected international scientific journals, memberships at
editorial boards of international scientific journals, participation at international research
projects, such as 5th and 6th Frame Project of EU, and a number of others. Departments of
immunology at the 1st and the 2nd Medical Faculty, Charles University, were approved by
Federation of Clinical Immunology Societies and became official centres of this particular
network. Investigators of this proposal also participate at the training of PhD students. Several
students successfully finished their PhD thesis. All members of the current research team
participated at the previous research, financially supported by the Ministry of Education of the
Czech Republic from 1998 to 2004, and they thus could extend the previously investigated
projects.
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C5.
Specification of the research plan objective
The aim of this research proposal is the elucidation of ethiopathogenesis and the improvement
of diagnosis and therapy of immunopathologic states. In accordance with the experimental
orientation of participating research teams, the research objectives are presented separately for
individual subprojects and divided into basic groups of immunological disorders: primary
immunodeficiencies, secondary immunodeficiencies, organ specific autoimmune diseases,
systemic autoimmune diseases, allergies, tumor immunology, infectious immunity, and
immunology of reproduction. A detailed description of all the investigated projects is
included in sections C5a-h. This section contains a brief summary of the whole research
proposal. As its integral part, the proposed project focuses on active involvement and training
of PhD studentshaving potentials to successfully finish their PhD studies and continue in
independent research activities.
a) The aim of the research in the field of primary immunodeficiencies is the elucidation
of the molecular basis of selected primary immunodeficiency diseases, evaluation of
the role of disease modifying genes, and the genotype-phenotype correlation. In the
field of management of patients with PID, we want to improve current therapeutical
protocols and adapt them to correspond to patients´ individual needs. The principal
investigator of this project is Assoc. Prof. Anna Šedivá, M.D., Ph.D., Charles
University, 2nd Medical Faculty.
b) The aim of the research in the field of secondary immunodeficiencies is the study of
pathogenesis of damaging immune reaction on the model of cardiovascular surgery.
Elucidation of the time course of the events at the molecular level (expression of
adhesive molecules, production of cytokines..) will help to define rational therapeutic
and preventive strategies that could restrict the extent of the inflammatory damage and
the degree of the postoperative secondary immunodeficiency. The principal
investigator of this project is Prof. Dr. Jan Krejsek, Ph.D., Charles University, Medical
Faculty Hradec Kralove.
c) The aim of the research in the field of organ specific autoimmune diseases is the
elucidation of the role of regulatory T lymphocytes in the pathogenesis of autoimmune
polyglandular syndromes and clarification of the principle of isohormonal therapy.
Role of cytotoxic T lymphocytes and other immunocompetent cells in the
pathogenesis of the axon damage will be studied on the model of the organ specific
autoimmunity affecting central nervous system – multiple sclerosis. Results will be
correlated with the stage and type of the disease with the aim to adjust a very
expensive immunotherapy of this disease on the individual basis. The principal
investigator of this project is Assoc. Prof. Ivan Šterzl, M.D. Ph.D., Charles University,
1st Medical Faculty.
d) The aim of the research in the field of systemic autoimmune diseases is the study of
patients producing defined autoantibodies in correlation with their genetic background
and with the response to therapy. Several groups of patients will be included: 1)
patients with rheumatoid arthritis producing autoantibodies against cyclic citrulinized
peptide (anti-CCP), 2) patients with polymyositis and dermatomyositis producing
autoantibodies against Jo-1 antigen and 3) patients with systemic lupus erythematosus
producing autoantibodies against ribosomal protein P. As the clinical relevance and
response to therapy according to the genetic background the presence of certain
antibody is not known, therapeutic consequences in selected groups of patients with
above mentioned diagnosis will be correlated with patients having different genetic
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e)
f)
g)
h)
i)
and autoantibody characteristics. The principal investigator of this project is Prof. Jiří
Vencovský, M.D., Ph.D., Charles University, 1st Medical Faculty
The aim of the research in the field of anaphylaxis is the study of pathogenesis of
acute allergic reaction (anaphylaxis), study of activation of effector cells and the study
of clinical significance of their experimental testing. The influence of the genetic
factors on the sensibilization and the development of anaphylaxis will also be tested.
The principal investigator of this project is Dr. J. Kučera, Charles University, 3rd
Medical Faculty
The aim of the research in the field of allergic disorders is the determination of the
significance of cellular and humoral inflammatory markers in the induced sputum of
patients with bronchial asthma or allergic rhinitis with regard to the course and
severity of the disease. The principal investigator of this project is Assoc. Prof. Petr
Panzner, M.D., Ph.D., Charles University, Medical Faculty Plzen
The aim of the research in the field of tumor immunology is the use of dendritic cellbased vaccines in the treatment of tumor diseases at the stage of minimal residual
disease. Research will be aimed at the understanding of the physiology of dendritic
cells, optimalization of antigen preparation and on the possibilities of minimal residual
disease detection. Project concentrates on the acute lymphoblastic leukemia of
childhood and on the ovarian cancer. The principal investigator of this project is
Radek Špíšek, M.D., Ph.D., Charles University, 1st Medical Faculty.
The aim of the research in the field of immunology of infectious diseases is the
optimalization of the early diagnosis of chronic neuroinfections (lyme disease,
bacterial meningitis) and the follow-up of the consequences of the chronic infection on
the development of immunopathologic states - secondary immunodeficiencies and
autoimmune disorders. This project is also focused on the improvement of diagnosis
and treatment of pulmory infections in patients with cystic fibrosis. The principal
investigator of this project is Pavel Pícha, Ph.D., Charles University, 2nd Medical
Faculty
The aim of the research in the field of immunology of reproduction is
 the evaluation of neuro-endocrine-immune parameters and immunologic
reactivity in pairs with the impairment of fertility in the corresponding
biological fluids.
 isolation of antibody isotypes and antigens from germinal cells and observation
of the course of affinity before, during and after the treatment.
 identification of dominant antigenic fraction responsible for the immune
reaction of immunocompetent cells.
 Problems of HIV infection in the reproductive period and its correlation with
the incidence of aborts will also be investigated. The principal investigator of
this project is Assoc. Prof. Z. Ulčová-Gallová, DrSc., Charles University,
Medical Faculty Plzen
C5a) Primary immunodeficiencies (Šedivá)
Primary immunodeficiencies represent a group of immune system disorders. Primary
immunodeficiencies are well defined units caused by disorders in genes that code different
parts of immune system. Primary immunodeficiencies are further divided into humoral,
cellular, combined, phagocytic disorders a complement deficiencies. In terms of research
intention we will work with primary immunodeficiencies as a whole being respectful of
characteristics of each group. There is strong tradition in taking care for patients with
primary immunodeficiencies in Institute of Immunology. This care is complex and covers
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diagnostic procedures, follow up and therapy of patients with primary immunodeficiencies.
Within the group of primary immunodefieicnces we’ll specifically concentrate on the group
of patients with syndromediGeorge and on the most severe forms of other immune deficiency
disorders (Chapter Humoral and Cellular and combined immunodeficiencies).
Humoral immunodeficiencies
Most severe disease of this group of disorders is Bruton´s agammaglobulinemia. We perform
long term basic health care for patients with Bruton´s agammaglobulinemia. In the research
project of the search for the molecular cause and particular mutation in btk gene we cooperate with St.Anne University Hospital in Brno in molecular and prenatal diagnostics of
our patients..
Within the group of patients with Bruton agamaglobulinemia we indetified patients with
contiguous deletion syndrom. Their deletion covers also another genes except of btk gene.
With the support of research project we want to finish genomic and proteomic analysis btk
and next gene DDP- DDP is the gene for the mitochondrial protein. We co-operate with
Centre for Mitochondrial disorders in this question.
We follow other humoral immunodeficiencies and we diagnose varieties of diseases, cased
by different gene mutations. This is performed in our laboratory and in co-operation with
other European countries. Research project support will enable further activity in this field.
Cellular and combined immunodeficiencies.
Institute of
Immunology is
the goal for the patients
with the most severe
immunodeficiencies - severe combined immunodeficiency (SCID). This patients require
bone marrow transplantation(BMT) for therapy.
Co operation of Institute of Immunology and Hematooncologic clinic, which performs BMT
in children ,enables this therapy.
Different gene mutations and influence of other modification genes requires individual
approach in diagnostics of this diseases. Approximately 2 patients per year (occurence of
SCID 1:50 000 – 1:100 000) are diagnosed in co operation with foreign laboratories.
Genetic mutations in each patient gives the direction of this research . Molecular diagnostics
if this diseases couldn´t be possible in Czech republic without the financial support of these
research activities.
Cellular and combined immunodeficiencies represent the most severe cases. Such patients are
concentrated in the Institute of Immunology and are shared with the Clinic of
Oncohematology as they required bone marrow transplantation. The individual diagnosis of
each patient is the team work of several national and international departments. As such
diagnosis is the research, often associated with the necessity to establish new research
methods, we reserve psrt of the research funding for these individual cases. Typically we take
care about 1 to 2 children with SCID each year.
We further concentrate on the group of patients with diGeorge syndrome.
These patients immunodeficiency is the consequence of missing or hypoplasy of thymus on
the basis of an embryopathy. Extensive deletion on chromosome 22 is the background of
this genetic disorder.
Immunologists, genetists, cardio - surgeons constitute the diagnostic and therapeutic team for
care for diGeorge patients. The initial research activity in children with diGeorge syndrome
was supported by grant IGA MZČR. We want to continue in the research with a view to
pathogenesis of diGeorge syndrome, mainly function of thymus in development of cellular
immunity and tolerance.
Low numbers of T lymfocytes in the first 2-3 years of life and dynamics of their
development represent our main research interest. Usually these findings are transient . The
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cytokines IL 7 and receptor for IL 7 play important part in this process. We want to observe
these molecules in the peripherial lymphocyte populations and in the thymus. We use human
peroperative thymus tissue taken and provided by cardiosurgeons in the time of heart surgery,
with the parents informed consent. In the preliminary study we proved that not all patients
have got complete absence of thymus tissue. Some of them have structurally altered even
nearly normal thymic tissue. The detailed study of thymuses in syndrom DiGeroge was not
yetr published. Support of this Research project would make possible to exploit the
thymic structure and function in the Institute of Immunology.
Phagocytic deficiencies and Complement deficiencies
Phagocytic deficiencies and deficiencies of complement are quite rare and now we do n´ t
plane research activity in this group . Nevertheless we take complex care of the patients
reffered to our department.
Objectives for the Research project
Complex diagnostics and therapy of primary immune deficiencies
1. Molecular genetic diagnosis in individual patients
2. DiGeorge syndrome pathogenesis
C5b) Cardiac surgical operation as a model for assessment of influence of complex stress
on immune system (Krejsek)
Deregulated function of immune system which is the principle part of inflammatory
response have a great potential to disturb homeostasis of an individual. Deregulation of
immune system could be the reason for morbidity and mortality of patients after cardiosurgery
operation using both cardiopulmonary bypass or without bypass. It is possible to find the
evidences of temporally deminished immune response, particularly specific immunity during
operation and through early postoperation period. The pathophysiological mechanisms of this
transient suppression of immunity after large operations are not known. It is not clear if this
postsurgical suppresion of immune reactivity is responsible for increased risk of development
of infectious complications. On the other hand, it is clear that the depression of specific cellmediated immunity which is proven in preoperative period by skin tests using T-dependent
anamnestic antigens is correlated with increased morbidity and mortality of patients in
postoperative period. There are no preoperation, peroperation, and postoperation parameters
which could identify the patients at the risk of development of unwanted immunopathological
reaction during cardiac surgery. A lot of variables play the role in the changes of immune
parameters of patients during cardiac surgery. Majority of cardiac surgical operations of
adults are performed on patients suffering from long-lasting atherosclerosis. Despite a lot of
contradictory results, current paradigma of atherosclerosis is based on the recognition of
principal role of inflammation in this process. This inflammatory reaction localized in blood
vessels wall could be induced by various stimuli either non infectious origin (modified
lipoproteins) or infectious origin (chlamydia) or combination of both factors. The
inflammatory process leading to the atherosclerosis is determined by genetic predisposition
and individual immune reactivity. This process is predominantly influenced by activities of
TH1 and TH2 subsets of T cells. Individual TH1 and TH2 reactivity is determined by genetic
background which is substantially influenced by variable conditions of environment during
early period of postnatal development. The function of immunoregulatory subsets of T cells
are determined by innate immunity, first of all by cells processing and presenting antigens to
T cells. The massive release of endogeneous “danger pattern” during cardiosurgery operation
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is probably associated with the exposition of innate immunity to these stimuli. The release of
endogeneous “danger pattern” is caused by the exposition of cells and humoral components of
immune system to artificial surfaces of supporting devices, by damage of cells and tissues,
generation of reactive oxygen intermediates, and by infectious agents. There is the trend to
compartmentalisation of immune response. There are limited data concerning the participation
of myocardial and lung tissue in the modulation of immune reactivity of patients undergoing
cardiac surgery. Without any doubt apoptosis will play an important role during cardiac
surgical operation. Apoptosis is induced in cardiomyocytes, lung parenchyma cells,
endothelial cells and other cells of nonhematopoietic tissues by hypoxia, by the presence of
free oxygen and nitrogen radicals, and cytokines. The apoptosis will be induced also in cells
of immune system either as the result of inadequate stimulation (e.g. contact activation of
granulocytes) or as a part of physiological regulation of immune response (absence of context
of recognition is leading to anergy or apoptosis of T and B cells).
C5c) T-regulatory lymphocytes in isohormonal therapy of organ specific autoimmune
diseases (Šterzl)
Aim of the work:
Collection of patients with autoimmune endocrinopathies (diabetes mellitus type I, Addison’s
disease, autoimmune thyroiditis) and selection of the appropriate form and timing for the
application of the substituting hormone in the form of isohormonal therapy in the early phase
of the disease development. The influence of this isohormonal application on the
development / suppression of the autoimmune process will be monitored focusing on Tregulatory lymphocytes activation.
Immune mechanisms in autoimmune polyglandular syndrome type II and III
Aim of the work:
We will monitor the mechanisms of activation of autoimmune processes in adult patients with
APS type II and III and with polyglandular activation of autoimmunity (PAA) selected at the
Institute of Immunology and Microbiology, 1st Faculty of Medicine, Charles University
Prague. These mechanisms lead often to subclinical manifestations of autoimmune
endocrinopathies.
We will monitor the dynamics of effector and regulatory lymphocytes, autoantibodies,
complement component C1q, naïve and memory lymphocytes – CD45RO+/CD45RA+
comparing the groups of patients with APS type II, APS type III and PAA.
The main task is to find the differences leading to the risk of the full clinical development of
the disease and of the protective factors leading to PAA.
Neuroimmunoendocrine regulations in organ specific autoimmune diseases
Aim of the work:
Monitoring of patients with sclerosis multiplex (MS) in relation to the therapy and type of the
disease. According to recent studies, an important role in the lasting axon disconnection also
in the clinically quiescent period without attacks in progressive types of MS play CD8+
lymphocytes. Further monitored groups in relation to the therapy will be patients with
myasthenia gravis and polyneuropathies. The monitoring of the patients will be performed
also in the relation to the stress, occurring not only due to the infection. The effort will
concentrate on disclosing of minute changes in the immune system accompanying the
disturbance of the neuroendocrine homeostasis not manifested by clinical impairment and
therefore not treated by clinical intervention. This disbalance can lead to the activation of
CD8+ lymphocytes and to subsequent axon damage in MS and probably also in some types of
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polyneuropathies.
The main task is to increase the cost effectiveness of therapy of patients with the
neuroimmunological diseases.
C5d) Genetic basics of autoantibody production in connective tissue diseases
(Vencovský)
Connective tissue diseases (CTD) are clinically characterized by the affection of different
organs with frequent manifestations in musculoskeletal system. It is a heterogeneous group of
diseases which is characterized by involvement of blood vessels and connective tissue as well
as by the presence of different autoantibodies. The important feature is the overlapping of
clinical and immunological abnormalities. CTD are mostly chronic, disabling diseases, but
relatively frequently also manifesting as severe subacute or acute illness with fatal outcome.
The total frequency of CTDs is slightly above 3 percent in the population and often young
persons are affected. The most severe CTDs are fortunately not so frequent and the prevalence
of these forms is about 1 percent. CTDs comprise rheumatoid arthritis (RA), systemic lupus
erythematosus (SLE), polymyositis and dermatomyositis (PM/DM), Sjögren´s syndrom (SS),
systemic scleroderma (SSc), mixed connective tissue disease (MCTD), overlap syndromes,
vasculitides, antiphospholipid syndrom (APS) and other more rare diseases.
The etiology of CTDs is largely not known. In a majority of patients various autoimmune
manifestations can be found, which are most probably responsible for the disease
development. Several factors may play a role, such as age, genetic background, hormonal
imbalance, abnormalities in immune regulations, infection, toxic and chemical compounds, or
drugs.
Genetic predisposition is polygenic, which means that several genes share the susceptibility to
disease. The most frequent association is described for HLA genes. The other probable
predisposing genes involve those that encode for cytokines, cytokine inhibitors, proteinases,
apoptotic proteins, immunoglobulins, T cell receptors etc. Autoantibodies, such as rheumatoid
factors, antibodies to CCP (cyclic citrullinated peptide), nuclear antigens, phospholipids,
endothelial antigens, cytoplasm of neutrophil leucocytes and to many other proteins, play the
important role in the pathogenesis of CTDs.
Genetic predisposition to autoantibody formation is not completely known. Some facts point
to the association with HLA molecules. This can be explained by the ability of multiple HLA
molecules to present (auto)antigens with various efficacy. But these associations cannot be
found every time, and also most of the healthy persons, who possess the same HLA
molecules, do not form pathogenic antibodies. Autoantibodies can be found more frequently
between family members of CTD patients, however, without necessarily being affected by the
disease. On the other hand, autoimmune diseases are more frequent between members of the
families, but the type of the disease may differ. This suggests general susceptibility to
autoimmune reactions.
We have only minimal information about the possible relationship between the autoantibody
formation and zygosity in HLA molecules. The density of pertinent HLA molecules may have
an effect on the antigen presentation and stimulation to autoantibody production. Little
information is also available about the polymorphism of other genes, such as immunoglobulin
or anticytokine genes. It is known that IL-4 and IL-10 can modulate antibody production, but
the relationship between their polymorphic variants to autoantibody production is only
partially explored. The relationship between cytokine levels in the peripheral blood, allele of
the cytokine gene, and formation of autoantibodies is also largely unknown.
The prognosis of patients with autoantibodies is usually worse than in patients without them.
We have recently shown that patients with anti-CCP antibodies and those with higher levels
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have more severe, erosive course of RA. A majority of systemic diseases has a variable course
and at the onset of the disease it is difficult predict the further development. It is therefore
important to have some parameters that can help in prognosis. The current possibilities are
clearly insufficient. This is closely associated with therapeutic approach to the patients. Two
facts are here important:
1) Because the etiology is unknown and pathogenesis is known only partially, the most
frequent therapeutic approach is non-specific immunosuppression. This is associated
with frequent side effects, which often depend on the drug dose. Glucocorticoid
induced osteoporosis or myopathy are good examples. The morbidity of a patient then
is a combination of the disease and treatment side effects. Occasionally, the
pharmacological toxic effects may be the significant contributors to the disability. If
we can predict with sufficient certainty the further course of the disease, we should be
able to dose the treatment appropriately and treat aggressively in patients with poor
prognostic factors, or more carefully in those with more favorable information. In the
former case we can try to prevent irreversible changes caused by the disease, in the
latter those changes caused by the unnecessary treatment.
2) The current possibilities of the treatment have broadened recently to the targeted
interference with basic molecular mechanisms. The biological treatment is effective in
about 70% of patients with RA. However, economic aspects have to be taken into
consideration besides the burden of the disease. Therefore those patients who have
higher diseases activity and particularly poor prognostic factors should be treated with
these drugs.
The aim of the proposed research is to find the relationship between genetic background in
CTDs and the ability to form autoantibodies. In the broader context then to relate these facts
to their prognostic significance and to the applied treatment.
C5e) Pathogenesis of the acute allergic reaction (anaphylaxis), risk factors for
anaphylaxis and its clinical usefulness. The influence of genetic factors on the
sensibilisation and anaphylaxis manifestation (Kučera)
The goal of the project is the identification of patients with high risk of anaphylactic reaction,
dominantly triggered by the hypersensitivity reaction Type I towards allergens from
Hymenoptera venom, with severe systemic manifestation (cardiovascular – hypotension,
shock, and respiratory symptoms). One of the major goals is to find risk factors or
mechanisms able to separate individuals with the sensitization with the low risk of the future
clinical responsiveness from individuals with sensitization and high risk of anaphylaxis
relapse, particular the severe reaction. One of these factors can be clinically silent mast cell
proliferation. Other resources (references) reported the prevalence of silent forms of
mastocytosis in patients with severe forms of anaphylaxis to be about 30 %, and the clinical
approach to those patients is different. In vitro studies showed antigen processing is different
according to the presence of particular MHC Class II alleles, some high risk alleles in the
relationship to the exposed allergen are described (wasp venom allergen or peanut proteins).
Part of this project will be coordinated with the multicentric European Academy of Allergy
Clinical Immunology study, directed by F. Rueff, focused to mastocytosis and venom
allergy, side effects risk factor identification during venom immunotherapy and failure of
venom immunotherapy.
Mechanisms and the efficacy of immunotherapy of anaphylaxis and allergy diseases –
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regulatory T cells and basophil reactivity.
Allergen immunotherapy is a causal, specific immunosuppressive treatment with the efficacy
ranged between 65 – 95 % according to the allergen used and the disease. Very good model is
venom induced anaphylaxis in monosensitized individuals, when the suppression of clinical
reactivity due to impairment of sensibilisation and changes of effector cells can bee estimated
by the standardized exposure test. In our project, the population of regulatory cells and
effector cells (peripheral blood basophil leucocytes) will be studied by measuring the
expression of membrane markers and cytokine production. The final goal will be the
identification of one or more laboratory markers for the future risk of clinical reactivity
estimation in patients already treated by venom immunotherapy, another words the individual
approach to patients which can bring higher effectivity of the therapy together with the
decrease of total cots for this long-term venom immunotherapy
C5f) Detection of cellular and humoral markers of inflammation in induced sputum
samples in patients with bronchial asthma and allergic rhinitis. (Panzner)
In effort to find the best methods used to assess airway inflammation in research and clinical
practice, induced sputum is one of the more referenced.
Irreversible structural changes in the airways caused by chronic inflammation have been
observed in mild asthma (beginning stage). Antiinflammatory optimal pharmacotherapy with
minimal or none adverse effects should be provided as soon as possible.
In atopic asthma the inflammatory process seems to be promoted by Th2 cytokines produced
by T cells (IL-4, IL-5, IL-13).
IL-4 stimulates B cells to produce IgE against aeroallergens and can induce VCAM-1
endothelial expression, eosinophils can better adhere to endothelial barriers.
IL-5 stimulates eosinophils differentiation and activation in bone marrow, has chemotactic
activity for eosinophils, has an impact on growing and survival rate of these cells, has
negative role in their apoptosis.
IL-13 has also ability to induce IgE production.
IgE molecules are binding on mastocytes Fc receptors in bronchial mucosa. After
sensitization, repeated contact with allergens can cause degranulation of mast cells, performed
mediators (histamine, tryptase) are released, leukotrienes, prostaglandines, tromboxanes and
PAF are produced in early phase of allergic reaction during 10-30 minutes. Histamine causes
bronchoconstriction and vasodilatation, increase vascular leakage and mucus hypersecretion.
Tryptase makes smooth muscle more sensitive to histamine. In the early phase of allergic
reaction, metabolites of arachidonic acid released from lung mastocytes make vasodilatation
and increase airway hyperresponsiveness (PGD2), LTC4, LTD4 and LTE4 increase vascular
permeability, mucus hypersecretion, bronchoconstriction and eosinophil chemotaxis, LTB4
chemotaxis of neutrophils. Activated mast cells are (as well as Th2 lymphocytes) the source
of cytokines IL-4, IL-5, IL-13. Mast cells play a role in late allergic reaction during 3-8 hours:
cell infiltration, especially eosinophils, less basophils and neutrophils, to the place of contact
with allergens. By this way Th2 answer is potentiated, allergic inflammation is persisting.
Expression of adhesion molecules on the endothelial cells and eotaxin production promote
eosinophilic infiltration in the late phase of the allergic reaction. The result is accumulation of
activated eosinophils in bronchial epithelium. Eosinophil is the main effector cell in the late
phase of allergic reaction. Basic proteins (ECP, EPX, MBP, EPO) released from eosinophil
cytoplasmatic granules cause denudation and disruption of the bronchial epithelium and
mucous hypersecretion. MBP cause depression of parasympaticus tension. Activated
eosinophils are hypodense and after degranulation are EG2+. We can identify them in
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bronchial biopsy, BAL or induced sputum from patients with asthma. Eosinophils are also the
source of prostaglandines, PAF and leukotrienes (LTC4, LTD4, LTE4) causing airway
hyperresponsiveness and edema. The result of long presence of activated eosinophils in tissue
is airway wall remodeling: increased collagen deposition in the basement membrane, mucus
gland hyperplasia, smooth muscle hypertrophy, vascular proliferation, loss of elastic fibers.
Bronchial epithelium plays a key role in pathogenesis of asthma, initiates development of
inflammation and bronchial remodeling (endothelins and TGF-β production stimulate
fibroblast to collagen hyperproduction).
Even during corticotherapy in asthma we can find markers of airway inflammation. The main
cells of airway inflammation - eosinophils (in lumen) and mastocytes (in epithelium) - are
corresponding with the degree of bronchial hyperresponsiveness in asthmatics treated by
inhaled corticosteroids.
Eosinophilic airway inflammation and epithelial damage can occur in patients with asthma,
airway inflammation persists even in those subjects, who are receiving inhaled corticosteroids
and who have normal lung function and good symptomatic control of their disease. Sputum
eosinophils and epithelial cells are significantly higher in patients with than without asthma.
Sputum compared with BAL showed a higher number of neutrophils, CD4+ and CD19+
lymphocytes, fewer macrophages and markedly higher levels of eosinophil cationic protein
(ECP), tryptase and fibrinogen. The findings in sputum differed from blood findings in
proportions of CD4+ and CD8+ T-cells, lower CD8+ cells and a higher CD4+/CD8+.
In induced sputum obtained from asthmatics and chronic bronchitis patients, the levels of
elastase were significantly increased and correlated with the percentage of neutrophils.
Airway inflammation in asthma and chronic bronchitis is associated with high levels of active
elastase.
In patients with mild asthma and animal dander allergy (Fel d1, Can f1) a significantly
decreased number of circulating eosinophils, increased bronchial sensitivity to methacholine,
increased sputum eosinophils (EPO) and myeloperoxidase (MPO) after animal allergen
exposure were observed.
Inflammatory cells are present not only in the airways of patients with asthma but also in
airways of patients with seasonal allergic rhinitis even without natural exposure. Basophils in
sputum of patients with asthma are present even during clinical remission. The presence of
bronchial responsiveness is associated with an increase in the number of eosinophils and
metachromatic cells (mast cells and basophils). Eosinophils, as well as mast cells, contribute
to bronchial responsiveness, not only in asthma but also in seasonal allergic rhinitis.
Although some non- asthmatic subjects with allergic rhinitis exhibit airway
hyperresponsiveness and increased diurnal peak expiratory flow (PEF) variation, little is
known about the critical features that determine these physiological alternations.
Eosinophilic inflammation may be present in subjects with allergic rhinitis and airway
hyperresponsiveness even when there are no symptoms of asthma. This could indicate that
bronchial eosinophilia is insufficient to cause asthmatic symptoms.
Apoptotic eosinophils are detectable in induced sputum of allergic patients. Apoptosis could
be an important mechanism in the control of acute eosinophilic inflammation in patients with
seasonal allergic rhinitis exposed to the sensitizing antigens. It appears that the apoptotic
mechanism could be less effective in controlling tissue eosinophilia in asthmatic patients with
chronic eosinophilic inflammation.
Patients with seasonal allergic rhinitis may develop bronchial hyperresponsiveness during the
active disease period. Eosinophil activation may occur in the bronchial mucosa and may be
reflected by increased sputum levels of ECP, especially when ECP binding proteins such as
α-2-macroglobulin pass through the lamina propria and across the epithelium into the airway
lumen.
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Histamine induced exudation of α-2-macroglobulin was associated with increased sputum
levels of ECP exclusively during the pollen season.
C5g) Dendritic cell-based vaccines in the immunotherapy of tumor diseases (Špíšek)
Dendritic cells (DC) represent the most efficient antigen presenting cells (APC). DC are the
only cells capable to induce primary immune reaction and to activate naïve T lymphocytes.
They circulate in the very small numbers in the peripheral blood and the difficulties with their
separation for the long time impeded their research. The study of DC has recently been
enabled by the description of protocols allowing for the generation of large numbers of DC
from peripheral blood monocytes in the presence of Granulocyte-monocyte-colony
stimulating factor (GM-CSF) and inerleukine-4 (IL-4). DC differentiate in the bone marrow
and they then migrate to the peripheral tissues where they exist in the immature state.
Immature DC express set of different molecules playing an important role in the process of
capturing of the surrounding antigenic particles (viruses, bacteria, infected, tumor and dead
cells). In their immature state, DC thus present, on their surface, the whole array of antigens
that corresponds to the composition of their environment. However, they are not equipped for
the efficient activation of T lymphocytes. This is a key mechanism for the homeostasis of
organism. Activation of the immune reaction in the steady state would increase the risk of the
development of autoreactive T and B lymphocytes and thus would increase the risk of an
autoimmune disease. Another scenario is represented by the situation when the organism is
confronted with the process that interferes with its normal function or is even dangerous for
its further existence. Danger could be represented by the number of stimuli. Classically, it is
the presence of a pathogen or large numbers of cells dying by necrotic death. Detection of
these specific signals starts the process that leads to the dramatic changes in the phenotype
and function of DC and that is called maturation. During the process of maturation DC lose
the ability to capture antigens, they highly increase the expression of the costimulatory
molecules (CD80, CD86), MHC class I. and II. molecules, and maturation specific molecules
(CD83). They further express, on their surface, a chemokine receptor CCR7. This molecule is
a receptor for the chemokine MIP-3, produced in the lymph nodes. Consequently, mature
DC migrated to the lymph nodes where it efficiently presents antigens to the antigen specific
T lymphocytes. DC at this stage also produce number of cytokines that further influence the
type of developing T cells. IL-12 is one of the most important ones.
Current therapeutic protocols used in oncology (surgery, chemotherapy, radiotherapy) are
relatively very efficient on their potency to reduce the number of tumor cells. The major
problem for the number of malignancies is the persistence of a small number of tumor cells
resistant to chemotherapeutics. This state is termed minimal residual disease (MRD) and there
is a relation between the level of MRD and the prognosis of a patient. Surviving tumor cells
give, later in the course of the disease, rise to the new population of cells resistant to therapy.
Immune system is able to eradicate tumor cells as demonstrated by the cases of spontaneous
remissions in malignant melanoma and in renal adenocarcinoma. Activation of an antitumor
immune response has several stages: 1) recognition of tumor antigens by immature DC, 2)
phagocytosis, processing and presentation of tumor antigens by DC, 3) activation and
proliferation of antigen specific T lymphocytes, 4) migration of effector T lymphocytes to the
tumor site and specific elimination of tumor cells. In each step, tumor cell can escape from
tumor surveillance. Tumor cells express antigens that are not present on normal cells. There is
a growing number of described tumor-specific or tumor-associated antigens (TSA, TAA,
respectively). Experiments in vitro have shown that if these antigens are presented to the
naïve T cells by mature DC they induce specific immune response. One of the reasons, why
tumor cells do not induce an immune reaction is the fact that they can function as the APC.
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As mentioned above, only professional APC can provide all signals necessary for the
activation of T lymphocytes. As tumor cell does not express the costimulatory molecules,
antigen-specific T lymphocyte is, upon encounter with tumor cell, driven into the state of
anergy, i.e. inability to proliferate and exert its cytotoxic function.
In accordance with the above-mentioned data, it appears that DC-based immunotherapy could
represent and elegant and efficient approach that could lead, in cooperation with other
treatment modalities, to the elimination of MRD. First clinical trials using DC pulsed with
tumor antigens showed very promising results in patients with end-stage malignant melanoma
and renal carcinoma. At the Institute of Immunology, we have studied DC biology and their
potential use in immunotherapy since 1996. Main interest of our research team concentrates
on hematological malignancies, especially acute lymphoblastic leukemia of the childhood
(ALL), and ovarian cancer. Results of our studies were published in a number of papers as
summarized in the list of literature. For the period of this research proposal we would like to
concentrate on the following aspects:
1) Definition of the optimal method for the generation of mature DC
2) Detail analysis of DC generated from peripheral blood monocytes of patients after
chemotherapy
3) Definition of a protocol for the preparation of the suitable form of tumor antigen from
leukemic blasts and the cells of ovarian carcinoma
4) Proposal of a Phase I, Phase II protocol for the clinical immunotherapeutic trial in
selected tumor diseases.
C5h) The immune response in some infections of the central nervous system in relation
to the proof of specific DNA and antigen respectively (Pícha)
CNS inflammations are a serious medical problem especially in the relation to a diagnostic,
treatment and following morbidity and mortality rate. The early and specific diagnostic
improves possibilities of a specific and symptomatic treatment. Moreover, in some cases of
neuroinfections the infection itself trigger immunopathological mechanisms which influence
the opening infection. Besides this, the induced immune changes are able to open a new
disease with immune etiopathogenesis that can continue without the infection agent. Detailed
information concerning of early diagnostic of the infective agent and immunopathological
reactions are an inevitable condition for more intensive study of the problem.
Lyme borreliosis is an infection known for tendency to spreading, multiorgan progress
and low invasivity of the pathogen. Spirochete infection usually progresses without symptoms
or as self-limited disease. Causative agent can persist in the organism resulting in chronic
organ inflammation. The main factor of persistence is previous degenerative organ changes.
However, the additional factors – mainly immune – will be necessary to find out. During last
years reduces a frequency of generalised forms of LB in clinical practise. Nevertheless, the
number of treated patients is relative high. Therefore it is necessary to develop a specific
diagnostic of LB, especially the direct methods of the pathogen proof. An antibodies based
diagnostic can have uncertain relation to the actual illness. DNA proof is practically the only
choice among the direct diagnostic methods due to high sensitivity because the body fluid
concentration of spirochete is usually low. Till now there don’t exist widely accepted routine
DNA kits and due to most of laboratory workers use “in-house” methods. These methods it is
necessary to design regarding to a specific disease form and body fluid. Results of DNA tests
have to be compared with other diagnostic tools – especially based on antibody detection.
The aim of this part of the project is the study of specific antibody production in nerve forms
of Lyme borreliosis and optimisation of methods of the DNA detection – looking at the PCR
itself and the primers too. An integral part of the problem is the clinical examination of the
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acute stage and recovery regarding to laboratory results.
Purulent meningitis are a persisting problem in infectious diseases. A key role in successful
choice of the antibiotic treatment plays its early starting. On the other hand it is limited by
early detection of the pathogen. Classic microbiological methods allow proving of the
causative agent till 24 hours. DNA techniques make possible the detection in a few hours.
Multi-stage techniques allow quick detection of the specific DNA in biological material by
combination of the broad-range and specific primers. Aim of the study in this part of the
project is the optimisation of detecting system based on PCR techniques.
Immunopathologic reactions with autoimmune features are known in CNS inflammations for
many decades. They interfere with a basic infective process in some cases of nervous
inflammations. The most frequently it is an isolated production of autoantibodies and/or
autoreactive clones of lymphocytes and disease results in recovery. In more complicated cases
disease needs an immunosupresive treatment. However, such treatment is in relative
controversy with necessity of the elimination of infective agent. From these reasons would
help a closer specification of the autoimmune processes to the more targeted treatment of
CNS inflammations. The aim of the study is preparing of methods for the detection of
autoantibodies and revealing theirs clinical meaning.
C5i) The aim of „Immunopathology of fertility failure“ (Ulčová-Gallová)
1. „Cells, antibodies and antigens of gametes resposible for decreased fertility“: to go on
studies of antibody activities in infertile couples, to concentrate on sperm antigenicity and
especially zona pellucida antigenicity, to determinate dominant epitopes and izolation of its
correspondent isotypes, to determine immunochemical properties of antigenic fractions and to
detect possibilities of desensibilisation or goal-directed immunisation on the contrary as
immunological planning of family.
2. Autoimmune endocrinopathology and fertility
.
The neuroendocrine axis in pairs with fertility disturbances will be followed up and
assessed both before and after conception and also after delivery. Our main emphasis will lay
on follow up of thyreopathies and on assesment of the presence of antibodies against thyroid
gland in various fertility disturbances mainly in endometriosis and polycystic ovary syndrom.
3.Thrombophilia and fertility
Genetic part of the project will be focused on the presence of thrombophilic mutations
(Factor V Leiden, G220210A mutation of prothrombin gene, C677T and A1298C mutations
of the MTHFR gene, mutations of PAI1 gene) in infertile women (e.g. APA I and APA II
syndromes) and especially in women treated with “megadoses” of glucocorticoids.
4.. HIV in the period of reproduction. The monitoring of viral loads in the seminal plasma
and sera in HIV-positive patiens before and after initiation of the Highly Active Antiretroviral
Therapy together with the evaluation of cellular and humoral immune parameters. In the
„Assisted reproduction programme“ in HIV-positive men of discordant couples (man HIV+,
woman HIV-) to introduce such a thrifty cleaning method (to get rid of HIV-1 and CD4+ T
lymphocytes) for seminal plasma and spermatozoa, which wouldn´t be functionaly disturbed
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C6.
Strategies and methods of the research plan solution
Strategies and methods are specified fro each section separately (C6a-h). It can be concluded
that for the support demanded for the whole research proposal is institutional. The aim is to
maintain the stability of the research at the medical faculties and to provide personal and
material for already existing research activities.
Strategies and methods include both the methods of basic and applied research. Training of
the young research fellows that would, in future, allow the continuity and development of the
research at university institutions forms an integral part of this project. Another part of the
strategy is the presentation of the results at the national and international conferences,
workshops, congresses and publications in Czech and international scientific journals. We
will also encourage the presentation of the results to the public in the form of popular articles,
lectures etc.
C6a)
We will continue to assemble and diagnose patients with primary immunodeficiencies. We
will continue to follow and to treat this patients. According to functional immunologic
tests we will target the diagnostic procedures to find molecular basis of the disease.
For the investigating of DiGeorge syndome patients we will use following methods:
Complex immunological investigation including immunophenotyping of
lymphocyte
subpopulations by flow cytometry especially double positive CD4+CD8+ T lymphocytes and
expression of IL 7 receptor CD 127
Investigation of IL 7 serum levels
Histological investigation of thymic tissue , staining of thymic lymphocyte subpopulations ,
following of
expression of IL 7 receptor in thymic tissue from children with diGeorge syndrome and
control group.
Confocal microscopy investigation of thymic tissue , passage of lymphocytes through thymic
tissue
Vaccination antibody response in childlren with diGeorge syndrome, postvaccination
antibody response development
Bcg vaccination response and its development
Now we follow aproximatelly 100 patients with the diagnosis of primary immune deficiency
in the Institue of Immunology, 39 of them with diGeorge syndrome. We expect aproximatelly
diagnosing of 5 new patients with diGeorge syndrome per year.
In the area of thymic tissue investigation we plane investigation of 10 samples in the year
2004, for consecutive years we will specify thymic tissue investigation according to our
results and accesibility of thymic tissue.
C6b)
Working hypothesis:
Cardiac surgical operation is complex stressful situation during which the integrity of
tissues is abolished by large operation and other harmful events such as hypothermia, hypoxia
and deminished blood perfusion. The overhelming deregulated activation of humoral and
cellular components of innate immunity is induced by both exposition to endogeneous
“danger patterns” and by contact activation in supporting devices. Other immunomodulating
factors are blood transfusion, administration of blood components, anesthesia, other
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medication including application of corticosteroids. The changes in mutual interactions
between immune system and neuroendocrine system are induced by these variables. The
exposition to undesired events during cardiosurgery is so extensive that compensatory
suppressive immune mechanisms are induced to prevent harmful activities of innate
immunity. As a consequence, cell mediated specific immunity is also depressed.
Postoperative immunodepression has to be limited to prevent the risk of infectious
complications in postoperative period.
Methods:
The scientific project supported by Ministry of Health, Czech Republic, entitled “An
attempt to decrease postoperative mortality by early diagnosis and treatment of viral
infections in patients undergoing cardiac surgery”, principal investigator dr. Pavel Kuneš,
Department of Cardiosurgery, University Hospital Hradec Králové was performed. We found
several significant changes in a representative group of 99 patients.
a) There are changes of cell mediated immunity. There is leukocytosis associated with
relative and absolute lymphopenia. There is the significant decrease in the number of
total T cells and subpopulation of helper inducer CD4+ T cells. The number of CD8+
cytotoxic suppressor T cells is increased. T cells reveal the signs of activation early
after surgery. There is increase in relative and absolute number of B cells. Immediately
after operation, there is significant increase in the number of NK cells which is
followed by their depletion in later period.
b) The level of humoral components of immunity and inflammation is changed during
and after cardiac surgery. The level of IgG, IgE and IgM isotypes of immunoglobulins
is decreased together with decreased level of selected components of complement
system. Serum level of components of complement system is increased in later period.
The similar trends are see for acute phase proteins. The significant activation of
immune system during operation is reflected by significantly increased level of 2
microglobuline. In contrast to decreased number of T cells during and after operation
there is the increase in the level of IL-2 and soluble form of IL-2 receptor.
Proinflammatory conditions are probably regulated by increased level of
proinfammatory cytokine IL-6. The serum level of soluble adhesion molecules
sICAM-1, sE-selectin and sVCAM-1 is already increased in patients undergoing
cardiac surgery before operation. This finding could implicate undergoing
inflammatory process associated with atherosclerosis in these patients. The elevated
level of soluble adhesion molecules is deminished during operation. The serum level of
soluble adhesion molecules reached previous level shortly after operation.
Aims:
We will focus on following parameters:
a) We will detect functional parameters of T cell system by measurement of proliferation
capacity of T cells after mitogenic stimulation in vitro by flow cytometry and cell cycle
analysis.
We will follow the level of selected cytokines skewed to functionally different subsets
of TH1 and TH2 T cells by immunofluorescence and flow cytometry, and by ELISPOT
technique. We will measure the level of CD40 ligand, IL10 and interferon gamma in
cell culture supernatants.
b) We will determine functional parameters of cellular components of innate immunity.
We will measure the oxidative burst of phagocytic cells by flow cytometry. The
expression of activation markers expressed on monocytes will be determined by
immunofluorescence and flow cytometry. The level of sCD14 and LBP molecule,
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neopterin and procalcitonin in serum, IL-12 and IL-18 in serum and cell culture
supernatants, will be determined.
c) Selected markers of apoptosis will be detected.
The apoptotic cells in blood and cell cultures will be identified by annexin test. The
overall rate of apoptosis will be assessed by the measurement of sApo/Fas.
Group of patients:
The patients will be recruited from patients surgically treated at the Department of
Cardiosurgery, University Hospital, Hradec Králové. The approval of local Ethical Commitee
together with informed written consent of patient will be obtained. The group of patients will
be split into two subgroups. The first group of patients will be operated with a help of
cardiopulmonary bypass. The second group of patients will be operated without
cardiopulmonary bypass. Another subdivision of patients will be based on their heart illness.
The group of patients suffering from ischemic heart disease and group of patients surgically
treated for valve defects will be delineated. All patients will be further subgrouped according
to the function of heart to patients with heart failure and patients with compensated heart
diseases. The samples of both arterial and venous blood will be collected before the
introduction to anesthesia, and after removal of aortal clamp. Only samples of venous blood
will be examined the first postoperative day, and the 4th, 7th postoperative day. The samples
collected from coronary sinus and lung vein will be obtained, eventually, during surgical
operation. No samples will be collected from coronary sinus and lung vein in patients who
will be surgically treated without cardiopulmonary bypass. Clinical parameters, variables
associated with cardiac surgical operation and clinical course in postoperative period will be
evaluated. These parameters will be corelated with parameters of immune system.
C6c)
T-regulatory lymphocytes in isohormonal therapy of organ specific autoimmune
diseases.
Activation of T-regulatory lymphocytes leads to the blockade of effector Th and Tc
lymphocytes. Treg lymphocytes are activated by presenting of the antigen produced by target
organs (hormones) by non-professional antigen presenting cells of low affinity that can be
reached by administering of the antigen e.g. through the gastrointestinal tract (most preferably
nasally). This mechanism is suitable for the breach of the development of the clinical
manifestations of the organ specific autoimmune damage.
Immune mechanisms in autoimmune polyglandular syndrome type II and III.
Autoimmune polyglandular syndromes (APS) represent a group of clinical manifestations
mainly taking part by the form of organ specific autoimmune disorders. Some secondary
conditions of these manifestations can be represented also by the humoral part of the damage
by the cytotoxic autoantibodies or by antibodies mediated by antireceptors. Many of these
various, mutually dependent immune mechanisms that have not yet been clarified. Different
APS types have different genetic predisposition – the greatest differences are between APS
type I and APS type II and III. Unlike APS type I, which is a homogeneous disease caused by
the mutation of AIRE gene, APS type II and III are of polygenous character and the model of
immune mechanisms for these diseases is more complicated. Epigenetical mechanisms of the
development of these diseases (unstable metals, infection) leading to different antigen
expression (e.g. 21-hydroxylase, 17-hydroxylase and other different cytochromes) will be
monitored. Besides of APS type II and III, also the group of the so-called polyglandular
activation of autoimmunity (PAA) belong to the group of the polyglandular autoimmune
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diseases due to the similarity in genetic predisposition. In this group the full clinical damage
of the target organ is not developed at all and the process of damage proceeds only on the
subclinical level, the full clinical damage is reached only at a late age, if ever.
Neuroimmunoendocrine regulations in organ specific autoimmune diseases
In the last ten years, the discipline of neuroimmunology has been growing very rapidly,
leading to a reevaluation of the hypotheses about the pathogenesis of many neurological
diseases. Our intention is therefore to prove the theoretical findings and findings acquired on
mice models in praxis. The monitoring of immunological parameters in patients with
neuroimmunological diseases is important for the diagnosis only partially, but very important
for the estimate of the disease development and suitable therapy. The changes in the
immunological parameters after beginning of the therapy of MS by different remedies
(interferon beta, Copaxone, combined immunosuppressive therapy, high doses of
immunoglobulins), where it is very difficult to choose the suitable strategy individually and
the clinical evaluation of the treatment benefit can be evaluated only after several months. The
immunosuppressive therapy is used also for other neuroimmunological impairments. The
basic method will be the immunophenotyping of peripheral blood and liquor by means of
flow cytometry – e.g., chemokine receptors on CD4+T-lymphocytes, determination of
CD8+CD28+ T-lymphocytes and intracellular determination of cytokines in CD4+ and CD8+
cells. The group under study will comprise of patients with neuroimmunological disease
attending to the Institute of Immunology and Microbiology of 1st Medical Faculty and
General Faculty Hospital, in cooperation with the Neurological Clinics of 1st Medical Faculty
and General Faculty Hospital.
C6d)
In the group of patients with RA those with anti-CCP antibodies will be identified and
compared with patients without these antibodies.
Similarly, patients with PM/DM with anti-Jo-1 antibodies (against histidyl-tRNA synthetase)
and with SLE and anti-ribosomal P proteins antibodies will be selected. The comparison will
be made with patients with different autoantibodies or with those without any defined
antibodies. HLA DR will be typed, and in RA also subtyping for the “shared epitope” alleles
will be performed. The polymorphism in IL-4 and IL-10 genes will be investigated and
related to present antibodies and clinical picture. Also the IL-1 locus will be considered,
where the polymorphism in IL-1 alfa and IL-1 beta genes will be related to the disease
presentation. All the polymorphisms and autoantibody presence will be compared with longterm disease course and response to treatment.
Methods
All patients will be investigated using standard questionnaires and the activity and damage
determined. In the case of RA it is DAS28, articular index 66/68, and HAQ; in the case of
PM/DM it is the newly developed tools MITAX, MYOACT, MYODAM and MDI; in the
case of SLE it is BILAG and SLICC/ACR. The quality of life and functional impairment will
be assessed also using the HAQ and SF-36. In PM/DM patients the functional index for
muscle endurance and muscle test scoring muscle weakness will also be used.
Autoantibodies will be determined with indirect immunofluorescence on Hep-2 cells, with
counterimmunoelectrophoresis (CIE) and with Western Blotting (WB). The extracts for CIE
will be prepared from calf thymus and from bovine spleen and tested against the panel of
standard sera. Extracts for WB will be made from HeLa cells in 3 ways – nuclear extract,
cytoplasmic extract and extract prepared in hypertonic conditions. These tests will serve for
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anti-Jo-1 and anti-P protein detection. In some cases the InnoLia methods will be used as
well.
The anti-CCP antibodies will be determined using the second generation ELISA.
For the longitudinal follow up of the antibody levels the ELISA test will be employed.
Gene polymorphisms will be determined from DNA derived from peripheral blood cells using
nested PCR. HLA molecules will be determined with sequence specific primers. Cytokine
gene polymorphism will be performed by amplification of the gene and digestion of the
products with specific enzymes.
C6e)
Pathogenesis of the acute allergic reaction (anaphylaxis), risk factors for anaphylaxis
and its clinical usefulness. The influence of genetic factors on the sensibilisation and
anaphylaxis manifestation.
Patients with the history of anaphylactic reaction will be examined and identified in the
Center for Hymenoptera venom allergy, on the basis of the clinical picture. Thos
phenotypically selected patients will be tested for the sensitization, by the quantification of
IgE, IgG4 specific for allergen, for the presence of elevated blood unstimulated tryptase
(UNICAP Pharmacia) and the presence of DRB1 (DRB1*07) a DQB1 HLA alleles. Genotype
frequencies will be compared with the population with low clinical reactivity and the normal
population. The DNA typisation will be performed by PCR with sequential specific primers
(genomic DNA from peripheral blood lymphocytes). This characterisation will help to more
precise the estimation of the likelihood of future anaphylactic reaction, it can help to discover
the group of patients with the indolent mastocytosis (those patients will be subsequently
tested on the cooperating haematology department)). This characterisation will subsequently
allow more precise indication of allergen immunotherapy and will be the leading aspect of
antigen dosage (therapeutic range of the maintenance dosis can be several times higher than
the commonly recommended dosis of 100 ug of venom extract for patients with the lower risk
of anaphylaxis relapse), it further can allow the estimation of the risk of side effects and the
duration of the treatment. Finally, this approach will increase the effectiveness of the
treatment.
Mechanisms and the efficacy of immunotherapy of anaphylaxis and allergy diseases –
regulatory T cells and basophil reactivity.
Allergen immunotherapy influences the immunology reaction not only by the shift of Th2
cytokines to Th1, but predominantly by the support of production of anti-inflammatory
cytokines (IL-10 a TGF-beta), originally from T regulatory and Th3 cells. Effector cells –
mastocytes and peripheral blood basophils are regulated by these T cells, but at the same time
they regulate T cells, dominantly by the IL-4 influence. The Treg cell phenotype
(CD4+CD25+), FcgammaRII (CD32) and complement receptors on T cells will be tested
during venom immunotherapy. The response of basophils will be tested by the expression of
ectonukleotid pyrophosphatase/fosfodiesterase 3 (CD203c) as a specific marker for basophils
and by the CD63 expression (protein from membrane vacuoles) on peripheral blood basophils
after the in vitro stimulation by relevant antigen. This reactivity will be related to the clinical
reactivity (field or challenge test), to the occurrence of side effects and the allergen dosage.
The long term efficacy after the end of the treatment will be followed similarly.
C6f)
The value of different cellular and humoral parameters from induced sputum will be
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evaluated for their potential practical use for assessing of the course of disease and
optimal subsequent treatment.
Three groups of patients (about 50 subjects each) will be followed up. Smaller groups of
subjects could bring greater statistical error at final evaluation.
1st group will be composed from the subjects with positive skin prick test with pollen
allergens or with positive specific serum IgE, examination being not older then 1 year, with
manifestation of allergic rhinitis or conjunctivitis during the pollen season, without asthma
symptoms.
2nd group: subjects with pollinosis with positive skin prick tests or positive specific IgE
antibodies to pollen allergens with asthma symptoms, without inhaled corticosterioid
treatment. Asthma will be diagnosed on the basis of functional tests being not older then 1
year (obstructive curve, decrease PEF, bronchial responsiveness with positive
bronchoconstriction with significant reversibility of FEV1 >-12% from baseline after
inhalation of short acting β- 2 agonist).
3rd group: subjects with negative skin prick test to common aeroallergens, without allergy or
asthma symptoms.
All subjects will be non-smokers or ex-smokers at least last 6 months, sputum will be induced
out of the period of respiratory infection. Clinical status will be checked by a physician
(documentation, questionnaire: character and relevance of the main symptoms, period and
length of allergy and asthma symptoms, contemporary therapy).
All included patients will be examined three times: 2 months before pollen season, during the
season when they have allergy or asthma symptoms and 2 months after the season in the
period without symptoms.
Sputum will be induced by inhalation of hypertonic saline aerosol (starting with 3% NaCl and
increasing to 4 % and 5% NaCl during the procedure) by an ultrasonic nebulizer (De Vilbiss
2000) after premedication with Ventolin 200 µg and processed by standard way (Pizzichini et
al.). Spirometry curve will be checked before and after the induction.
In all acquired samples of sputum total cell count and viability, expression of cell markers and
soluble markers will be assessed.
Cytospines will be used for different cell count and immunocytochemistry staining (MBP+,
elastase+ and CD68+ cells).
In supernatant IL-4, IL-5, IL-10 and IL-13 will be assessed.
Cytometric detection of EG2 and final choice of other markers (CD 45/14, CD14/HLADR,
CD 9/CD69) will be probably correlated with research development.
The results will be compared in respect to time period of sampling and among different
groups of persons, analyzed by using T-test and Wilcoxson test and published.
C6g)
The groups of the patients studied in our project are represented by children treated for ALL
and patients with the ovarian cancer.
1) Definition of the optimal method for the generation of mature DC
Maturation of DC is a prerequisite, as only mature DC can activate immune response.
Maturation can be achieved by the stimuli mimicking the presence of infection
(proinflammatory cytokines, lipopolysacharide, double stranded RNA, bacterial
DNA). Each of these activators probably modulates DC function differently. We will
define an optimal activator for the use in the immunotherapy.
2) Detail analysis of DC generated from peripheral blood monocytes of patients after
chemotherapy.
Candidates of immunotherapy approaches are patients with MRD who have already
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underwent the reduction of tumor mass by another treatment modalities. Some of them
i.e. chemotherapy or radiotherapy are likely to influence the function of DC. In this
part of the project, we will investigate, whether DC generated after anticancer therapy
have phenotypic and functional characteristics comparable to DC from healthy donors.
3) Definition of a protocol for the preparation of the suitable form of tumor antigen from
leukemic blasts and the cells of ovarian carcinoma
An approach for the tumor diseases, in which TSA are not defined, is the use of
apoptotic tumor cells or tumor lysat as a source of tumor antigens. Immune response
induced in this case is polyclonal and the incidence of escape variants of tumor cells is
less probable. A frequent objection to this strategy is the potential danger of inducing
an autoimmune disease. However, it appears that the breakdown of mechanisms
ensuring tolerance of self tissues is more demanding than the induction of antitumor
response. These findings correlate with our results, which found no sings of
autoreactive cytotoxic T lymphocytes.
4) Proposal of a Phase I, Phase II protocol for the clinical immunotherapeutic trial in
selected tumor diseases.
Administration of DC to healthy volunteers was well tolerated without any serious
adverse effects. Based on these results Food and drug administration (FDA) approved
the DC-based immunotherapy as a therapeutic strategy in humans. Currently, there are
several studies on different tumor diseases in Europe and USA. During this project we
will define a clinical protocol for those tumor disease, where it will be possible to
define an optimal form and quantity of tumor antigen used for DC pulsing. Protocol
will be approved by Ethical committee of Teaching Hospital Motol and the Research
Council of the Ministry of health.
Methods
Ad 1) We will compare morphology, phenotype and functional characteristics of generated
DC. For the phenotypic analysis, we will determine surface and intracellular expression of
antigen presentation associated molecules (HLA-ABC, HLA-DR), maturation-related
molecules (CD83, DC-LAMP), adhesion and costimulatory molecules and relevant
chemokine receptors. For the assessment of DC functional properties we will concentrate on:
 ability to induce T cell proliferation (thymidine incorporation, CFSE labelling)
 Th polarization (measurement of cytokine production by ELISA and flow cytometry)
 Capacity to engulf tumor cells (flow cytometry, confocal microscopy)
 Induction of cytotoxic T cells (ELISPOT, tests of cytotoxicity)
Ad 2) Blood samples from patients treated for ALL will be drawn at the end of the induction
therapy and DC will be generated from peripheral blood monocytes. In patients with ovarian
cancer, peripheral blood samples will be taken before the treatment and after chemotherapy.
Characteristics of DC will be determined by the same methods as in 1. Blood samples will be
taken during regular check-ups after obtaining an informed consent. MRD level will be
regularly determined as a part of complementary research projects. We plan to enroll 10
patients for the group of ALL and 15 patients for the group of ovarian cancers. DC generated
from the blood of healthy blood donors will be used as a control.
Ad 3) We plan to use either apoptotic tumor cells or defined epitopes derived from the
chromosomal translocations for the induction of cytotoxic T lymphocytes. Cytotoxic T
lymphocytes will be generated by repeated stimulation with antigen pulsed DC. Generated
cytotoxic T cells will be compared for their capacity to lyse tumor cells.
Ad 4) In the course of this project, we will define a clinical immunotherapeutic protocol based
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on our results. Generation of DC vaccine for the use in vivo has to follow very strength
criteria and all the procedures and used reagents must be of GMP quality. Thus, for this
project and for other protocols of cellular therapy investigated at the Charles University, we
plan to build a Cell therapy unit, which will fulfill the EU criteria. Part of the investments for
the equipment for this unit is included in this research proposal.
C6h)
In patients with acute and chronic manifestations of Lyme borreliosis it is intended to
examine the production of antiborrelial antibodies in blood and CSF. Specific antibodies will
be detected in CSF by antibody index CSF/serum. In CSF will be examined oligoclonal bands
too and in well-founded cases will be verified theirs antigen specificity (Borrelia
Burgdorferi.s.l.). During the study will be the antibody production compared with spirochete
DNA presence using PCR. The PCR method will be optimised by the selection of suitable
primers and others methodical steps. The main purpose in this part it is introducing of new
primers working specifically in acute and chronic forms of Lyme disease. An important
component of the study is deepening of the PCR DNA detection in urine, which is worldwide
connected with hesitation. But, clinical profit would be substantial. Laboratory methods will
be correlated with a clinical progress of patients in a minimal period of 6 months. This latency
is necessary especially in Lyme borreliosis, where only under this condition is possible to
assess the result of treatment. In this part we suppose inclusion of 120-150 patients during all
period.
In purulent meningitis it is intended to test step by step the DNA detection system consisting
of broad-range and specific primers. In the case of positive results would be the more suitable
species-specific primers of the main pathogens in purulent meningitis tested. PCR results
would by correlated with cultivation techniques and with specific antibodies.
The third main objective of the study is more detailed study on autoimmune reactions in CNS
inflammations. Methodical tolls will be especially the detection of autoantibodies against
nerve antigens (MBP,GFAP) in body fluids (serum, CSF) by ELISA and immunoblotting.
Results will be correlated with clinical findings and treatment.
C6i)
Informed infertile couples with written consent will be examined by routine gynaecologic,
immunologic, endocrinologic and genetic methods. According to predominat cause of
infertility, the next examination will be aimed genetically or “according to “infectional rules”.
Methods:
1. microagglutinating tests (tray micragglutination test, mixed antiimmunoglobulin reaction
tests)-for detection of sperm antibodies in serum, in seminal plasma, in ovulatory cervical
mucus, in peritoneal fluid, in follicular fluid.
Passive haemmagglutination and ELISA- antibodies against zona pellucida
ELISA- panel for detection of antiphospholipid antibodies against cardiolipin, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, pho-acid, β2-glycoptotein I, annexin V.
Extraction-solubilization methods including SDS PAGE, gel chromatography,
immunoblotting, immunofluorescent method will be used for isolation of sperm antigens, and
for isolation of zona pellucida antigens. Laboratory equipment will serve for isolation of each
isotype of Ig to each antigenic fraction.
Immunochemical and immunohistological analysis will follow individual, particular antigenic
fractions of spermatozoa and zona pellucida in the process of desensibilization and active
immunization.
2. To follow up neuroendocrine axis we will monitor changes in sex hormones levels (day
3rd., 14th and 27th ) and assess thyroid function (TSH, free T4 and free T3) and anti thyroid
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gland antibodies using radioisotopic, chemiluminiscence and enzymatic ligand
(immunoanalytical) assays.
3. We will use ASA PCR, PCR with Hinf I for detection of Leiden mutation for factor V, G
20210 A , C677T.
4. Commercial kits will be used for detection of VL (viral load), flow cytometry (f.r. for CD4
and CD8 lymphocytes), active retroviral therapy and affin. chromatopgraphy.
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C7.
Time schedule of the research plan solution
None of the projects included in our proposal starts on the green land. Each proposal is the
continuation of the projects supported in past by previous research aims of the Ministry of
Education. We will apply the principle of continuous evaluation of the results reached during
the study and the publication of interim results. Contribution of each group to the will be
evaluated at the regular meetings at the end of the year. Based on the defined criteria
(publications, presentations, new diagnostic and therapeutic methods, number of PhD students
etc) the financial support for each year will be allocated according to the efficiency of each
team. Specific timetable of each project are detailed separately in sections C7a-i.
C7a)
See previous paragraph
Detection and classification of new patients into research groups will run simultaneously as
well as their diagnosing and therapy.
We expect to reach the number of approximately 60-70 patients with diGeorge syndrome
during duration of the Research project. So we can form unique group of children with this
disease.
Planed investigation will be performed simultaneously according to the methods introduced
in the previous paragraphs
We plane the investigation of 10 samples of thymic tissue in the year 2004, for consecutive
years aproccimatelly 5 new samples per year.
C7b)
The objectives of this scientific project could be reached with diagnostic procedures which are
available at our department. At least 50 patients will be enrolled into this study every year.
Approximately 400 patients are surgically treated at our Department of Cardiosurgery every
year, so as recruitment of 50 patients could be without limitation. It is very likely that
objectives of this scientific project will be modified during realisation according to the
scientific results obtained in this field in the world. The progress in diagnostic methods,
technologies and improvements in cardiac surgical operations will be also reflected. The total
number of patients enrolled ensures detailed stratification of patients on the base of evaluated
clinical and laboratory parameters. We suppose that it could be possible to formulate general
conclusions at the end of this project.
C7c)
T-regulatory lymphocytes in isohormonal therapy of organ specific autoimmune
diseases.
In 2005, a group of patients with the above-mentioned autoimmune endocrinopathies will be
established and these patients will start to be substituted perorally or nasally by the substituted
hormone. In the following years a method of determination of the activity of Treg
lymphocytes will be worked out and other parameters of the autoimmune process –
autoantibodies, T effector lymphocytes, the ratio of the Th1/Th2 and Tc lymphocytes - will be
monitored.
Immune mechanisms in autoimmune polyglandular syndrome type II and III.
During the first year the selection of patients included in the study will be performed. During
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the following years the parameters distinguishing the immune mechanisms in the particular
groups will be monitored.
Neuroimmunoendocrine regulations in organ specific autoimmune diseases
The examination of patients will be performed throughout the whole duration of the project,
the new patients can be included in the study at any time, after every year an evaluation and
possibly also a change in the monitored parameters will be performed.
C7d)
In the first and second years the patients will be identified and investigated clinically. The
serological typing will be performed at the same time. In all patients the basic clinical
parameters will be determined as well as the aspects evaluating activity, damage and quality
of life. The activity will be assessed every 6 months, damage once a year. The longitudinal
variation of autoantibodies is not known and therefore these will be estimated in 3-months
intervals, so that we can answer the question about their fluctuation in serum. A possible
predictive value of this phenomenon will be determined. The cytokine levels will be detected
and related to the allele present in the genome and to the level of autoantibodies and their
fluctuation. In the subsequent years the DNA will be isolated and polymorphism of selected
genes will be determined. Patients will be long-term followed and the effect of therapy will be
evaluated in the subgroups.
C7e)
Pathogenesis of the acute allergic reaction (anaphylaxis), risk factors for anaphylaxis
and its clinical usefulness. The influence of genetic factors on the sensibilisation and
anaphylaxis manifestation.
The first group of patients with the appropriate clinical picture will be enrolled during 2005,
the risk factors identification, particularly to the long term efficacy will be performed.
Subsequent patient enrollment with the recent history will be continual
Mechanisms and the efficacy of immunotherapy of anaphylaxis and allergy diseases –
regulatory T cells and basophil reactivity.
The methodology of the laboratory part will be started and validated for the diagnosis in
2005; methods will be tested as prognostic markers next years.
C7f)
We expect to enrol approximately 50 patients yearly. In the first two years of the investigation
we will have an effort to choose patients suitable for this follow up in accordance with the
form and degree of affection. Then in the course of whole 5 years the patients will be enrolled
progressively to the separate groups in accordance with above- mentioned criteria.
In next years we will follow dynamics of markers of inflammation in induced sputum in
connection with undergoing therapy in these patients (allergen immunotherapy,
antihistamines, topical corticosteroids treatment).
C7g)
During the first 3 years we will enroll newly diagnosed patients with ALL or ovarian cancer.
In the in vitro experiments we will define the optimal conditions for the isolation of tumor
antigens, DC generation, tumor antigens presentation and for the induction of DC maturation.
An integral part of this highly clinical research will be the study of DC physiology that would
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elucidate the mechanisms that make these cells the key component of the immune system.
At the same time we will work on the proposal of a protocol of DC-based immunotherapy in
patients with ALL and ovarian cancer and we will prepare the documentation for the Cell
therapy unit. In the second half of the duration of this project, we plan to initiate Phase I and
Phase II of the clinical immunotherapeutic trial based on the DC vaccination. Enrolled
patients will be continuously monitored for the parameters of antitumor immunity and for
their clinical status.
C7h)
According to the specification of the study we suppose selection and testing of the primers in
Lyme borreliosis during first two years. The primers reactive in acute and chronic disease will
be tested. Verifying of the PCR methods for urine is intended to perform. During followed
years will be PCR used in a clinical practice.
In the PCR study in purulent meningitis it is planed a testing of the broad-range primers in
first 2 years and later, in the second 2 years a quality of species-specific primers will be
tested. A clinical conclusion will be done in the end.
The study of autoimmune reactions will be performed continuously in both of two abovementioned groups and, additionally, in next suitable cases of neuroinfections. In the end of
study will be performed laboratory and clinical estimation. In all variants of the study it will
be necessary to act according to the actually finance resources, which will be the limiting
factor of investigation.
C7i)
Prospective study includes systematic examination of couples with fertility failure according
to harmonogramme of neuro-endocrine-immune axis, and genetic examination. Individual
approach will be done according to cause of infertility. We expect huge increasing of infertile
couples from 17 IVF centers (see strategies 1-4).
2005- 2007: collection of sperm cells after swim-up (donors, fertile healthy men)
collection of unused no-fertilized oocytes
collection of serum, seminal plasma, of HIV positive patients
flow cytometry to detection of T lymphocytes in positive patients
PCR in genetic analysis
ELISA and immunofluorescency in detailed hormonal profiles
2007 -2008 extraction-solubilization methods for antigen isolations of sperm, zona pellucida,
gel chromatography, electrophoresis, immunoblottings, immunohistochemical and
immunohistological analysis
2007 -2010: early publications of our results, affin. chromatography and next ways for
“purification” of sperm and seminal plasma from HIV in patients being ready for IVF.
C8.
Envisaged results of the research plan solution (including the specification of its
character, subject matter and time schedule of its envisaged application)
Results will be published continuously and they concern both basic research
(ethiopathogenesis of immunopathologic states) and applied research (improving of diagnosis
and therapy of these disorders). They will be published or introduced into clinical practice as
soon as they will be reviewed in the journal or by the appropriate institutions. Individual
interim results are specified in sections C8a-h together with their time schedule. Publicity and
presentation of the results of this research proposal will be regularly presented to the scientific
authorities. One of the regular forums for these presentations will be at the conferences of
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Czech society for alergology and clinical immunology. Each month one of the working
groups will organize a section focused on its research activities and results. Results will be
also presented at the national and international conferences.
C8a)
Research activity in the field primary immunodeficiencies is necessary for keeping in
touch with science of European Union . Europe has traditionally very high level of
knowledge, overtaking U.S.A. in primary immunodeficiency field.
The research is necessary for precise diagnosis of this diseases on the molecular genetic
level.
In the future we expect the detailed diagnosis of proteins, their modified expression and
function.
The research project will allow us to be up to date in comparison with other countries and to
continue with the co- operation within the European Society for Immunodeficiencies ESID
We expect reaching priority results in the field of pathogenesis of immunodeficiency in
immune system
with missing thymus .
C8b)
The original data which enriched our understanding to the influence of complex stress on
immune system during cardiac surgical operation will be obtained in this study. The first
objective is to determine to role of activation of innate immunity and the participation of TH1
and TH2 immunoregulatory subsets in the operation induced pathophysiological reaction. The
second objective of this study is to determine the participation of heart and lung compartment
on this process. The third objective of this study should be the identification of selected
parameters of immune system which could identify the patients treated by cardiac surgical
operation who could be at the risk of overwhelming complications during operation and in
post operation period.
C8c)
T-regulatory lymphocytes in isohormonal therapy of organ specific autoimmune
diseases.
Improvement of the therapeutic model in patients with autoimmune endocrinopathies and
suppression of the development of autoimmune endocrinopathies in the early stages of the
disease.
Immune mechanisms in autoimmune polyglandular syndrome type II and III.
Selection of diagnostic and predictive markers distinguishing the groups with full clinical
manifestation of the disease from patients with only subclinical signs of the disease. The
achieved results will be exploited for the therapeutic intervention in patients with
polyglandular autoimmune disease that will not allow the development of the full irreversible
clinical symptoms based on the destruction of the target cells.
Neuroimmunoendocrine regulations in organ specific autoimmune diseases
Improvement of the effectiveness of the therapeutic procedures in patients with
neuroimmunological disease – selection of the appropriate remedy, modification of the
treatment during infection and stress.
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C8d)
The results of this project should answer the questions about the relationship between
autoantibodies and several disease aspects, such as the relations to the disease severity,
predictive capacity for further disease development and the ability to interfere with this
development by targeted therapy. The significance of autoantibody level fluctuation will be
determined and related to different disease aspects. Genetic background and the ability to
form specific autoantibodies will be established at the level of HLA and cytokine genes. The
significance of the gene polymorphism for the actual production of cytokines together with
the effect on autoantibody production will be assessed. All these facts will be correlated with
the clinical disease duration and with the response to treatment. In the end these facts will be
helpful in the prediction of disease course and will enable the introduction of targeted and
differential treatment, so that this is in started timely and in sufficient doses in the
prognostically unfavourable cases, and in only minimal required doses in those cases with
favourable prognostic parameters, so that toxic effects are avoided.
C8e)
Pathogenesis of the acute allergic reaction (anaphylaxis), risk factors for anaphylaxis
and its clinical usefulness. The influence of genetic factors on the sensibilisation and
anaphylaxis manifestation.
The improvement of the disease diagnosis and dominantly the identification of patients with
high need of intensive approach to allergen immunotherapy. On the other hand, identification
of individuals with the low risk, when the therapy can be less intensive
Mechanisms and the efficacy of immunotherapy of anaphylaxis and allergy diseases –
regulatory T cells and basophil reactivity.
The higher understanding of the mechanism of allergen immunotherapy leads to more precise
therapy indication during the project, the individual treatment will bring the benefit for some
patients during the course of therapy. The individual results for the patients is the significant
decrease of the morbidity, eventually mortality risk, from the public health point of view, the
final effect is the rationalization of the treatment (the reduction of expenses in patients with
the low risk of anaphylaxis relapse).
C8f)
Examination of sputum samples could be held as one of the valuable non-invasive method to
monitor airway inflammation, to follow up the development of disease and potentially to start
earlier with adequate treatment.
We expect, that we find in sputum of asthmatic subjects allergic inflammation with
corresponding laboratory changes.
In pollinotics, inflammatory markers will be probably also present, the intensity of
inflammation will be probably mild compared to asthmatics. During the pollen season
inflammatory changes will be more pronounced accordant with clinical symptomatology, in
control group they will be probably missing.
The literature references are quite controversial. Inflammation in the airways does not
probably persist long time after the pollen season, but inflammatory cells are present in the
airways of patients with seasonal allergic rhinitis.
In our study we want to verify the presence of inflammatory changes during the pollen season
and out of the season, assess the effect of treatment in patients with allergen immunotherapy.
Eosinophilic inflammation in the lower airways will be observed not only in sputum of
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asthmatics, but also in pollinotics without asthma symptoms.
There is a question for further investigation if these outcomes will affect the way and the
intensity of the treatment.
The finding, if markers of inflammation are present in pollinotics also before and after the
pollen season and if the markers of inflammation are present even in sputum of patients only
with symptoms of allergic rhinitis can be significant indicator for modification of the
treatment.
C8g)
Research in the field of immunotherapy by DC in tumor diseases and the initiation of clinical
trials has a great potential as a complementary treatment modality of tumor diseases.
Experimental parts of this project can contribute to the better knowledge of general
mechanisms of tumor immunity. Close connection with the projects of minimal residual
disease detection investigated at the 2nd Faculty of Medicine should in future lead to the
identification of patients in an increased risk of tumor relapse and initiate the treatment early
after the initial therapy by other modalities. We presume that the results obtained during this
project will be sufficiently novel to be published in respected scientific journals as for our
previous projects. They should form a base for the initiation of the first DC-based
immunotherapeutic trials in the Czech Republic.
Beside the publication activity, the main goal of this project is the application of this
promising approach into the clinical practice. Effective antitumor immunotherapy
administered at the stage of MRD could substantially contribute to the better prognosis of
patients with tumor diseases.
C8h)
We expect an improvement PCR technique for the diagnostic of acute and chronic forms of
LB. Results of PCR will be evaluated from the clinical point of view and compared with
specific immune reaction of the organism – the specific antibodies production in blood and
CSF. The more detailed information about oligosmyptomatic and atypical clinical forms of
LB could be expected due to PCR diagnostic – especially the clinical manifestations with low
production of antibodies or seronegative forms. The possibility of a dynamic examination of
PCR in urine would permit to observe the long-term elimination of the specific DNA from
organism. This way would allow obtain the maeningful marker of the treatment effectiveness
because neither the antibody tests, nor clinical symptoms provide sufficient information. In
the time of termination of the study it could be expected the routine laboratory and clinical
using of verified methods.
In purulent meningitis it is possible to expect the results especially in improving of early
diagnostic of the disease and related clinical consequences. We can expect routine clinically
using of PCR methods when results will be positive.
Proceeding study in the region of autoimmune nervous inflammation can bring more
information in parainfectiuos immunopathological reaction in relation to the particular tissue
antigens and autoantibodies. The conclusions could be expected in the end of the study.
C8i)
Envisaged results in “Immunopathology of fertility failure”
1.
to improve the quality of immunological cause of infertility or decreased fertility, the
utilization of antigenicity of each epitopes (spermatozoa, zona pellucida) gained from
extraction-solubilization methods for the process of active immunization and/or in the process
of desensibilization, or family planning.
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2.
3.
4.
2. to improve the diagnosis of infertility, to improve the hormonal treatment and the therapy
of autoimmune cause of infertility as thyropathie.
3. to find and to treat genetically diagnosed trombophilic diseases, and antiphospholipid
syndrome I-II
4. in discordant couples HIV positive (man HIV positive, woman HIV negative) being in
reproductive age, to base in praxis “purification” of seminal plasma and sperm cells from
HIV-1 and CD4 lymphocytes
All gained results will serve to qualification of diagnosis of infertility, improvement of
therapy. We expect the increasing of the Czech population , and EU population indeed
(today, Special consultation for reproductive immunology has examined : 7 couples from
Germany, 2 couples from Italy, 2 couples from France, 1 couple from Switzerland, 1 couple
from Gr.Britain, 12 couples from Russia, 1 couple from USA, 1 couple from Arab.Emirates,
11 couples from Ukraina, 1 couple from Japan, 4 couples from Vietnam, 3 couples from
China.
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D
D1.
Personnel guarantee
List of leading research employees according to names of the applicant
substantially and creatively participating in the solution of the research plan
together with the data on their age, envisaged major activities heading to the
fulfilment of research plan objective and envisaged working capacity expressed
as a working load in per cent
Surname
Name
Year of
birth
Bartůňková
Šedivá
Jiřina
Anna
1958
1955
Krejsek
Jan
1958
Šterzl
Ivan
1954
Vencovský
Jiří
1953
Kučera
Petr
1957
Panzner
Petr
1958
Špíšek
Radek
1975
Pícha
Dušan
1957
Ulčová-Gallová
Zdenka
1957
Dřevínek
Marečková
Hrdá
Pútová
Sedláček
Janda
Pavel
Helena
Pavlína
Ivana
Dalibor
Aleš
1975
1963
1971
1961
1957
1974
Liška
Martin
Mejstříková (PhD
Ester
student)
Rožková (PhD student ) Daniela
Principal activity
Working
load in %
50
50
1971
1977
coordinator of VZ
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experimetns
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
coordination
of
research,
evaluation of the results, design of
experiments
laboratory work
laboratory work
laboratory work
laboratory work
laboratory work
management of database, patient
follow-up
laboratory work
laboratory work
1979
laboratory work
50
44
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
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Surname
Tobiášová (PhD
student)
Seifertová (PhD
student)
Hölzelová (PhD
student)
Brázová (PhD student )
Virtová (PhD student)
Name
Year of
birth
Principal activity
Working
load in %
Zuzana
1978
laboratory work
100
Daniela
1977
laboratory work
100
Eliška
1972
management of PID database
100
Jitka
Martina
1977
1979
laboratory work
laboratory work
100
50
D2.
Structure of degrees of other members of the investigatory team together with the
number of persons, envisaged major activities aiming at the fulfilment of goals of
the research plan and envisaged working capacity in hours per year
Number of
Working
Degree group
Major activities
persons
capacity
Lab asistant
2
cytometry
4192
Lab asistant
1
cytometry
210
Lab asistant
1
laboratory work
210
Lab asistant
1
cell cultivation
629
Lab asistant
1
evaluation of results, statistical analysis
629
Lab asistant
1
laboratory work
2 096
Lab asistant
1
laboratory work
2096
Statistician
4
data collection
2516
Lab asistant
2
examination of genetic polymorphism
2516
Lab asistant
1
laboratory work
210
Data manager
2
data collection
420
Lab asistant
2
cytometry
4192
Lab asistant
1
cytometry
419
Lab asistant
3
laboratory work
4401
Lab asistant
4
laboratory work
1677
Lab asistant
2
imunogenetic testing
838
Lab asistant
2
laboratory work
419
Lab asistant
2
laboratory work
419
Lab asistant
2
laboratory work
419
Lab asistant
1
laboratory work
2096
Lab asistant
1
laboratory work
2096
Lab asistant
5
laboratory work
3145
Lab asistant
2
laboratory work
4192
Lab asistant
1
laboratory work
1048
Nurse
1
dealing with patients
419
Lab asistant
1
laboratory work
2096
Lab asistant
1
laboratory work
1048
Lab asistant
2
laboratory work
2096
Lab asistant
1
laboratory work
838
Lab asistant
1
collection of samples
210
Adminstrative
1
administrative work, laboratory work
2096
worker
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Adminstrative
worker
Adminstrative
worker
Adminstrative
worker
D3.
2
administrative work, laboratory work
419
1
administrative work
210
1
administrative work
210
Auxiliary personnel for the assurance of subsidiary activities necessary for the
solution of the research plan and envisaged working capacity in hours per year
Characteristic of a subsidiary activity
Washing of laboratory tools
D4.
Working
capacity
629
List of major implemented research and development applications of the
members of the investigatory team mentioned in D1 which refer to the issue of
the research plan, within the last five year
J.Bartůňková
Publications:
1. 1995-7 Systémové vaskulitidy a patogenetický význam protilátek proti cytoplasmě
neutrofilů, IGA MZ 3160-3, přidělené fin. prostř. Kč 890.000,- Ukončeno kat. A
2. 1998-2000 ANCA v dětském věku, IGA MZ 4553-3, přidělené fin. prostř. Kč 1
665.000,-ukončeno kat. A.
3. 2000-2002 Expozice křemíku a ANCA-asociované vaskulitidy, IGA MZ 6308-3,
přidělené fin. prostř. Kč 1 381.000,- Ukončeno kategorií A.
4. 1998-2000 Úloha adhezivních interakcí v imunitním systému. studie na
experimentálním zvířecím modelu myši s funkčně poškozenými selektinovými
ligandy. GA UK 177/98, přidělené fin. prostř. 670.000,-. Ukončeno kat. B.
5. 1997 Komplexní výuka klinické imunologie. FRVŠ 1375, přidělené fin. prostř.
221.000,6. 1999-2004 Komplexní imunologický program. Výzkumný záměr MŠ J13/98:
111300001,
7. Metodologie přípravy dendritických buněk, IGA NI 5669-3, 1999-2001, ukončeno kat.
A (spoluřešitel, řešitel dr. D. Pospíšilová), přidělené fin. prostředky 1 354 094 Kč.
8. Příprava individuálních nádorových vakcín na bázi dendritických buněk, IGA NI
7537-3, 2003-2005, vstupní hodnocení A.
9. BARTŮŇKOVÁ, Jiřina. Imunodeficience. Praha: Grada Publishing, 2002. 232 s.
10. HOŘEJŠÍ, Václav, BARTŮŇKOVÁ, Jiřina: Základy imunologie. Triton, Praha, 1998,
2. vydání 2002.
11. BARTŮŇKOVÁ, Jiřina. Primární imunodeficity a sekundární imunodeficity. In
Nouza, Martin and Nouza, Karel. Imunologie`98. Praha: Galén, s. 19-35.
12. BARTŮŇKOVÁ, Jiřina. Průtoková cytometrie v diagnostice defektů imunity. In
Eckschlager, Tomáš. Průtoková cytometrie v klinické praxi. Praha: Grada Publishing,
1999. s. 61-76.
13. BARTŮŇKOVÁ, Jiřina.Diagnostika a monitorování autoimunitních chorob. In
46
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Eckschlager, Tomáš. Průtoková cytometrie v klinické praxi. Praha: Grada Publishing,
1999. s. 80-86.
14. BARTŮŇKOVÁ, Jiřina. Využití průtokové cytometrie u transplantací. In
Eckschlager, Tomáš. Průtoková cytometrie v klinické praxi. Praha: Grada Publishing,
1999. s. 97-109.
15. BARTŮŇKOVÁ, Jiřina. Imunitní poruchy u pacientů se selháním ledvin. In Sulková,
Sylvie a kol. Hemodialýza. Praha: Maxdorf. Jesenius, 2000. s. 693-703.
16. ŠEDIVÁ, Anna,, KOLÁŘOVÁ, Ivana, , BARTŮŇKOVÁ, Jiřina. Antineutrophil
cytoplasmic antibodies in children. Eur J Pediatr, 1998, vol. 157, no.12, p. 987-991.
IF 1.318
17. ŠEDIVÁ, Anna, BARTŮŇKOVÁ, Jiřina, KOLÁŘOVÁ, Ivana, , HRUŠÁK, Ondřej,
VÁVROVÁ, Věra, MACEK, Milan jr., LOCKWOOD, Martin Ch., DUNN, Austyn C.
Antineutrophil Cytoplasmic Autoantibodies (ANCA) in Children with Cystic Fibrosis.
Journal of Autoimmunity, 1998, vol. 11, p. 185-190. IF 2.297
18. HRUŠÁK, Ondřej, TRKA, Jan, ZUNA, Jan, HOUSKOVÁ, Jana, BARTŮŇKOVÁ,
Jiřina. STARÝ, Jan. Aberrant expression of KOR-SA3544 antigen in childhood acute
lymphoblastic leukemia predicts TEL-AML1 negativity. Leukemia, 1998, vol. 12, p.
1064-1070. IF 3.163
19. TESAŘ, Vladimír, MAŠEK, Zdeněk,, RYCHLÍK, Ivan, MERTA, Vladimír,
BARTŮŇKOVÁ, Jiřina, STEJSKALOVÁ, Anna, ŽABKA, Jiří, JANATKOVÁ,
Ivana , FUČÍKOVÁ, Terezie, DOSTÁL, Ctibor, BEČVÁŘ, Radim.: Cytokines and
adhesion molecules in renal vasculitis and lupus nephritis. Nephrol Dial Transplant,
1998, vol.13, p. 1662-1667. IF 2.056
20. TESAŘ, Vladimír, JELÍNKOVÁ, Erna, MAŠEK, Zdeněk,, JIRSA, Milan jr.,
ŽABKA, Jiří, BARTŮŇKOVÁ, Jiřina, STEJSKALOVÁ, Anna, JANATKOVÁ,
Ivana, , ZIMA, Tomáš: Influence of Plasma Exchange on Serum Levels of Cytokines
and Adhesion Molecules in ANCA-Positive Renal Vasculitis. Blood Purification,
1998, vol. 16, p. 72-80. IF 1.251
21. DOSTÁL, Ctibor, TESAŘ, Vladimír, RYCHLÍK, Ivan, ŽABKA, Jiří, VENCOVSKÝ,
Jiří, BARTŮŇKOVÁ, Jiřina, STEJSKALOVÁ, Anna, TEGZOVÁ, Dana. Effect of 1
year cyclosporine A treatment on the activity and renal involvement of systemic lupus
erythematosus: a pilot study. Lupus, 1998; vol. 7, no. 1, p.29-36. IF 2,514
22. HOLÍKOVÁ, Zuzana, SMETANA, Karel Jr., BURCHERT, Maria, DVOŘÁNKOVÁ,
Barbora,., BOVIN, Nikolaj V., KLUBAL, Radek, BARTŮŇKOVÁ, Jiřina. LIU, FuTong, GABIUS, Hans-Joachim. Expression of galectin-3, galesctin-3-binding epitopes
and of Ga1/Ga1NAc-binding sites in keratinocytes of the adult human epidermis.
Electronic Journal of Pathology and Histology, April-June 1999, 992.
23. BARTŮŇKOVÁ, Jiřina. ŠEDIVÁ, Anna, VENCOVSKÝ, Jiří., TESAŘ, Vladimír.
Primary Sjögren`s syndrome in children and adolescents: Proposal for diagnostic
criteria. Clinical and Experimental Rheumatology, 1999, vol.17, p.381-386. IF 1.638
24. HRUŠÁK, Ondřej, TRKA, Jan, ZUNA, Jan, BARTŮŇKOVÁ, Jiřina, STARÝ, Jan:
Are we ready to curtail testing for TEL-AML1 fusion? Leukemia, 1999, vol. 13, no.6,
p.981-983 IF 3.163
25. SMETANA, Karel jr., HOLÍKOVÁ, Zuzana, KLUBAL, Radek, BOVIN, Nikolaj V.,
DVOŘÁNKOVÁ, Barbora, BARTŮŇKOVÁ, Jiřina, LIU, Fu-Tong, GABIUS, HansJoachim. Coexpression of binding sites for A(B) histo-blood group trisaccharides with
galectin-3 and Lag antigen in human Langerhans cells. J.Leukocyte Biology, 1999,
vol. 66, no.4, p.644-649. IF 4.262
26. ŠEDIVÁ, Anna, SMETANA, Karel jr., STEJSKAL, Josef, BARTŮŇKOVÁ, Jiřina,
LIU, Fu-Tong, BOVIN, Nikolaj V., GABIUS, Hans-Joachim. Binding sites for
47
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carrier-immunobilized carbohydrates in the kidney: implication for the pathogenesis of
Henoch-Schonlein purpura and/or IgA nephropathy. Nephrol.Dial.Transplant, 1999,
vol.14, p.2885-91.
IF 2.056
27. VENCOVSKÝ, Jiří, JAROŠOVÁ, Kateřina, MACHÁČEK, Stanislav,
STUDÝNKOVÁ, Jana, KAFKOVÁ, Jarmila, BARTŮŇKOVÁ, Jiřina, NĚMCOVÁ,
Dana, CHARVÁT, František. Cyclosporine A versus Methotrexate in the treatment of
polymyositis and dermatomyositis. Scand J Rheumatol., 2000, vol. 29, no. 2, p. 95102. IF 1.396
28. BARTŮŇKOVÁ, Jiřina, MALÝ, Petr, SMETANA, Karel, jr., ŠEDIVÁ, Anna,
KLUBAL, Radek, MAYEROVÁ, Dita, SEDLÁČEK, Aleš, ŠPLÍCHALOVÁ,
Veronika. Reduced phagocytic activity of polymorphonuclear leukocytes in (1,3)
Fucosyltransferase VII-deficient mice. Acta Pathol Microbiol Immunol Scand, 2000,
vol. 108, p. 409-416. IF 1.713
29. ŠEDIVÁ, Anna, VÁVROVÁ, Věra, BARTOŠOVÁ, Jana, POHUNEK, Petr,
BARTŮŇKOVÁ, Jiřina, MACEK, Milan Jr. Immune Aspects of Cystic Fibrosis.
Allergy Clinical Immunology International , 2001, vol. 13, no.2, p. 67-70.
30. ŠEDIVÁ, Anna, HOZA, Josef, NĚMCOVÁ, Dana, POSPÍŠILOVÁ, Dagmar,
BARTŮŇKOVÁ, Jiřina, VENCOVSKÝ, Jiří. Immunological investigation in children
with juvenile chronic arthritis. Med Sci Monit, 2001, vol. 7, no.1, p. 2-7.
31. BARTŮŇKOVÁ, Jiřina, KOLÁŘOVÁ, Ivana, ŠEDIVÁ, Anna, HOLZELOVÁ,
Eliška. Antineutrophil cystoplasmic antibodies (ANCA), anti-Saccharomyces
cerevisiae antibodies (ASCA) and specific IgE, to food allergens in children with
inflammatory bowel diseases. Clinical Immunology, 2002, vol. 102, no. 2, p.162-168.
IF 2,377
32. LUKÁŠ, Jaromír, SMETANA, Karel jr., PETROVICKÝ, Pavel, PALEČKOVÁ,
Věra, VACÍK, Jiří, DVOŘÁNKOVÁ, Barbora, BROŽ, Lubomír, POSPÍŠILOVÁ,
Dagmar, HOLÍKOVÁ, Zuzana, BARTŮŇKOVÁ, Jiřina. Biological properties of
copolymer of 2-Hydroxyethyl methacrylate with sulfopropyl methacrylate. Journal of
Materials Science: Materials in Medicine, 2001, vol. 12, p. 639-646. IF 0,701
33. POSPÍŠILOVÁ, Dagmar, BOROVIČKOVÁ, Jiřina, POLOUČKOVÁ, Andrea,
ŠPÍŠEK, Radek, ŠEDIVÁ, Anna, HRUŠÁK, Ondřej, STARÝ, Jan, BARTŮŇKOVÁ,
Jiřina. Generation of functional dendritic cells for the potential use in the therapy of
acute lymphoblastic leukemias. Cancer Immunology Immunotherapy 2002, vol. 51, p.
72-78. IF 2,820
34. Starý J., Sedláček P., Vodvářková Š., Gašová Z., Bartůňková J.: Development of
common variable immunodeficiency in a patient with Evans syndrome treated by
autologous stem cell transplantation. Pediatric Allergy and Immunology,
2003(14):334-337. IF 1,5
35. Šedivá A., Bartůňková J., Bartošová J., Jennette C., Falk R.J., Jethwa H.S.:
Antineutrophil cytoplasmic antibodies directed against bactericidal/permeability
increasing protein detected in children with cystic fibrosis inhibit neutrophil-mediated
killing of P.aeruginosa. Microb. Inf. 5,2003:27-30. IF 1.96
36. Bartůňková J., Tesař V., Šedivá A.: Diagnostic and pathogenetic role of antineutrophil
cytoplasmic antibodies. Clinical Immunology 10,2003: 73-82. IF 2.76
37. Cimaz R., Casadei A., Rose C., Bartůňková J., Šedivá A., Falcini F., Picco P.,
Taglietti M., Zulian F., Ten Cate R., Sztajnbok F.R., Voulgari P.V., Drosos A.A.:
Primary Sjögren syndrome in the paediatric age: a multicentre survey. Eur J Pediatr.
162,2003,10:661-665 IF 1.22
48
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A. Šedivá
1. Grant IGA 1999-2002: Imunitní systém u dětí se snydromem DiGeorge, hodnocení B.
2. Šedivá A., Bartůňková J., Litzman J., Zachová R.: Immune System in Children with
Di George Syndrome. VIII. Meeting of ESID, Rhodos. In: Molecular Immunology 35,
August 1998
3. Bartůňková J., Starý J., Kobylka P., Šedivá A., Pospíšilová D., Vávra V., Hrušák O.,
Friedrich W.: Hematopoetic Stem Cell Transplantation for Primary
Immunodeficiencies in the Czech Republic. VIII. Meeting of ESID, Rhodos. In:
Molecular Immunology 35, August 1998
4. Bartůňková J., Starý J., Kobylka P., Šedivá A., Pospíšilová D., Vávra V., Hrušák O.,
Friedrich W.: Hematopoetic Stem Cell Transplantation for Primary
Immunodeficiencies in the Czech Republic. VIII. Meeting of ESID, Rhodos. In:
Molecular Immunology 35, August 1998, 11-l12: 771
5. Šedivá A., Bartůňková J., Nevoral J.: Hepatitis B in a child with a deficiency of the
C4A component of complement. Third International Conference on Therapies for
Viral hepatitis, December 1999, Maui, USA. in Antiviral Therapy 1999; 4
(Supplement 4):abstract 109.
6. Richterová J., Bartůňková J., Šedivá A.: Deficit lektinu vázajícího manózu. Alergie 3,
1999:158-158
7. Šedivá A., Starý J., Hromadníková I., Skalická A., Ghio M.: Využití vyšetření
solubilních HLA molekul I. třídy u dětí při transplantaci kostní dřeně. Čas. lék. čes.,
139, 2000, 20:630-634
8. Bartůňková J., Malý P., Smetana K. Jr, Šedivá A., Klubal R., Mayerová D., Sedláček
A.,
9. Šplíchalová V.: Reduced phagocytic activity of polymorphonuclear leukocytes in
alpha(1,3)
10. fucosyltransferase VII-deficient mice. APMIS 108, 2000, 6: 409-416
11. Hadač J. Šedivá A., Asplund L., Smith E., Zeman J., Houštěk J.. Mohr-Tranebjaerg
(dystonia-deafness) syndrome and agammaglobulinemia. 4. kongres EPNS, BadenBaden. In: European Journal of Pediatric Neurology, 2001 vol 5, 5:A110
12. Šedivá A.: Vývoj imunitního systému v dětském věku, Medicína v praxi 3, 2001:4143
13. Kočárek E, Krutílková V, Puchmajerová A, Šedivá A, Bartůňková J, Němečková M,
Klein T, Zapletal R, Novotná K, Strnad M, Sálová M, Novotná D, Hlavovicová M,
Hejtmánková M, Malíková M, Maříková T, Baxová A, Simandlová M, Vejvalková Š,
Goetz P: Vztah genotypu a fenotypu u pacientů s mikrodelecí chromozomu 22q11.
Čs.pediat. 2001, 56,8:427-437
14. Šedivá A., Čiháková D., Lebl J.: Immunological findings in patients with autoimmune
polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) and their family
members: Are heterozygotes subclinically affected? J Pediatr Endocrinol Metab
2002,15:1491-6
15. Šedivá A., Bartůňková J., Zachová R., Hrušák O., Kočárek E., Novotná D., Novotná
K., Klein T.: Vývoj imunity u syndromu DiGeorge. Alergie 1, 2003: 8-13
49
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J. Krejsek
Grants:
1. Detekce reziduální choroby mnohočetného myelomu po vysokodávkové chemoterapii.
řešitel: MUDr. V. Maisnar, oddělení klinické hematologie. RČ IGA MZ ČR: 4535-3,
C, ukončen 2000
2. Konfigurace imunoblotu jako konfirmační sérologické metody v diagnostice Lymeské
boreliózy. řešitel: MUDr. K. Honegr, infekční klinika. RČ IGA MZ ČR: 4549-3,
ukončen 2000
3. Dlouhodobý vliv hemodialýzy a peritoneální dialýzy na imunitu nemocných
s chronickým selháním ledvin. řešitel: MUDr. P. Fixa, CSc., I. interní klinika. RČ IGA
MZ ČR: 4565-3, B, ukončen 2000
4. Stanovení funkční aktivity a fenotypu tumor infiltrujících lymfocytů u
nemocných s ovariálními tumory. řešitel: Mgr. Miroslava Toušková, ÚKIA. IGA MZ
ČR. NH/5196-3/99, B, ukončen 2001
5. Adoptivní imunoterapie jako součást kombinované léčby metastatického
postižení jater. řešitel: doc. MUDr. Bohuslav Melichar, Ph.D., II. interní klinika. IGA
MZ ČR. NI/4676-3/98, A, ukončen 2000
6. Adenomy hypofýzy kultivované in vitro: vliv somatostatinových analogů
a induktorů apoptózy. řešitel: MUDr. Jan Čáp, CSc., II. interní klinika. IGA MZ ČR.
NB/6172-3/00, projekt ukončen k 31.12.2002
7. Úloha metabolické, enoteliální a trombocytární dysfunkce v procesu
atherogenese. řešitel: prof. MUDr. Milan Bláha, CSc., OKH FN. IGA MZ ČR
NB/6549-3, projekt v řešení
8. Proteomové centrum pro studium intracelulárního parasitismu bakterií. nositel:
Vojenská lékařská akademie JEP, Hradec Králové. odp. řešitel: MUDr. Jiří Stulík,
CSc. Ostatní.VC LN00AO3, zahájeno v roce 2000, dosud trvá
Publications:
1. Pudil, R., Pidrman, V., Krejsek, J., Gregor, J., Tichý, M., Andrýs, C., Drahošová, M.:
Cytokines and adhesion molecules in the course of acute myocardial infarction. Clin.
Chim Acta 280(1-2), 1999, 127-134. IF 1.067
2. Krejsek, J., Žák, P., Toušková, M., Vokurková, D., Kodydková, K., Kvardová, A.,
Kopecký, O.:  T cell lymphoproliferation: a case report. Eur. J. Haematol., 2000, 64 (1),
68-70. IF 1.846
3. Tošner, J., Toušková, M., Melichar, B., Dítětová, I., Krejsek, J., Kopecký, O.: Phenotype
of tumor infiltrating lymphocytes (TIL) in patients with malignant ascites. Int. J. Gynecol.
Obstetrics, 2000; 70(Suppl. 1): 137-138. IF 0.376
4. Pudil, R., Krejsek, J., Pidrman, V., Gregor, J., Tichý, M., Bureš, J.: Inflammatory
response to acute myocardial infarction complicated by cardiogenic shock. J.
Submicroscop. Cytol. Pathol., 2000; 32(3): s.377. IF 0.687
5. Havlasová, J., Hernychová, L., Halada, P., Pellantová, V., Krejsek, J., Stulík, J., Macela,
A., Jungblut, P.R., Larsoon, P., Forsman, M.: Mapping of immunoreactive antigens of
Francisella tularensis live vaccine strain. Proteomics, 2002; 2: 857-867. Databáze I SCI
Journal IF za rok 2000 ani 2001 neuvádí. Rok 2002 ještě není vložen. Jedná se o nový
časopis. Je považován za hlavní světový časopis v oblasti proteomiky.
50
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Publications concerning the theme of the project:
6. Kuneš, P., Krejsek, J.: Změny v imunitní odpovědi po operacích s použitím mimotělního
oběhu: možná příčina rozvoje fatálních systémových infekcí vyvolaných oportunními
organismy a viry. Remedia - Klin. mikrobiol., 1, 1997, 10-12.
7. Kuneš, P., Žáček, P., Lonský, V., Krejsek, J.: Reperfuzní poškození v kardiochirurgii. IV.
část: Mimotělní oběh a celková zánětlivá odpověď organismu. Cor Vasa, 39, 1997, 340348.
8. Kuneš, P., Žáček, P., Lonský, V., Krejsek, J.: Reperfuzní poškození v kardiochirurgii. Cor
Vasa, 40(8), 1998, 399-407.
9. Kuneš, P., Žáček, P., Lonský, V., Krejsek, J.: Reperfuzní poškození v kardiochirurgii. Cor
Vasa, 41 (5), 1999, s. 231-240.
10. Kuneš, P., Krejsek, J.: Imunosupresivní cytokiny a zánětlivá odpověď v kardiochirurgii.
Cor Vasa, 42, 2000, 1, s. 47-54.
11. Kuneš, P., Krejsek, J.: Hojení ran z pohledu kardiologie a kardiochirurgie. Cor Vasa,
2000; 42(5): 244-250.
12. Kuneš, P., Krejsek, J.: CD4+ lymfopenie a pooperační imunosuprese v kardiochirurgii.
Časopis lékařů českých, 2000; 139(12): 361-368.
13. Kuneš, P., Krejsek, J.: Endotel a reakce obranného zánětu. Cor Vasa, 2001; 43(3): 138143.
14. Kuneš, P., Krejsek, J.: Ateroskleróza, imunitní odpověď a protein tepelného šoku
HSP65/60. Cor Vasa, 2001; 43(4): 205-212
15. Kuneš, P., Krejsek, J.: C-reaktivní protein a reakce akutní fáze. 1. část: CRP a bakteriální
infekce. Cor Vasa, 2002; 44(11): 467-475.
16. Kuneš, P., Krejsek, J.: C-reaktivní protein a reakce akutní fáze. 2. část: CRP a řízení
zánětlivé odpovědi. Cor Vasa, 2002; 44(12): 525-535.
I. Šterzl
Grants:
1. IZ/3419-3 Vliv těžkých kovů na vývoj únavového syndromu a autoimunních
endokrinopatií, IGA MZ ČR, 1996-1998
2. IZ/4205-3 Vliv cytokinů lymfocytových subpopulací TH1 a TH2 na vývoj
endokrinních autoimunitních onemocnění IGA MZ ČR, 1997-1999
3. NB/5394-4 Interakce neuroendokrinních a imunologických faktorů v mužském
reprodukčním systému" IGA MZ ČR 1999- 2002
4. 311/98/1036 Ovlivnění autoimunitních endokrinopatií cytokiny lymfocytárních
subpopulací GA ČR. 1998- 2000
5. ME 319 Diagnóza oftalmopatií, MŠMT 1997-1999
6. ME 375 Molekulární charakteristika autoimunitního polyglandulárního syndromu,
MŠMT 2000-2002
Publications:
1. Bártová, J., Procházková, J., Krátká, Z., Benetková, K., Venclíková, Z., Šterzl, I.
Dental amalgam as one of the risk facotrs in autoimmune diseases. Neuroendocrionol.
Lett., 24(1-2), 2003, 65-67.
2. Šterzl, I., Hampl, R., Hill, M., Hrdá, P., Matucha, P. Immunomodulatory cytokines in
51
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human seminal plasma correlate with immunomodulatory steroids. Steroids 68(9),
2003, 725-731.
3. Hrdá,P., Šterzl, I., Matucha, P. Cytokine levels in sera of patients with autoimmune
endocrinopathies. Physiological.Research. 52, 2003, 256-267.
4. Šterzl, I., Hrdá, P.,Matucha, P., Korioth, F., Kromminga, A. Porovnání autoimunitních
endokrinopatií v oblasti exprese HLA antigenů. Alergie 4, 2002, 281-284
5. Šterzl, I., Votruba, J., Matucha, P., Šterzl, J. Influence of Th1 a nd Th2 cytokines on
the primary and secondary immune response in vitro. In. From Proteomics to
Molecular Epidemiology: relevance of Autoantibodies (Report on the 6 th Dresden
Symposium on Autoantibodies held in Dresden on September 4-7.2002). K. Conrad,
M. Fritzler, M. Meurer, U. Sack, Y. Shoenfeld (eds.), Pabst Science Publishers,
Lengerich, Berlin, Riga, Rom, Viernheim, Wien, Zagreb, 2002, 248-249.
6. Hrdá, P., Korioth, F., Matucha, P., Šterzl, I., Kromminga, A. HLA association with
autoimmune endocrinopathies. In. From Proteomics to Molecular Epidemiology:
relevance of Autoantibodies (Report on the 6 th Dresden Symposium on
Autoantibodies held in Dresden on September 4-7.2002). K. Conrad, M. Fritzler, M.
Meurer, U. Sack, Y. Shoenfeld (eds.), Pabst Science Publishers, Lengerich, Berlin,
Riga, Rom, Viernheim, Wien, Zagreb, 2002, 248-249.
7. Pohanka, M., Hampl, R., Šterzl, I., Stárka, L. Steroid hormones in human semen with
a particular respect to dehydroepiandrosterone and its immunomodulatory metabolites.
Endocrine Regul. 36, 2002, 79-86.
8. Hrdá, P., Šterzl, I. Autoimunitní polyglandulární syndromy. Alergie, vol.3, 2001, 126129.
9. Veselý, D., Astl, J., Šterzl, I. Cytokiny u nádorů štítné žlázy. Alergie 3, 2001, 247252.
10. Martínek, J., Šterzl, I., Šterzl., J. Functional morphology of Thyroid gland "in vitro"
and "in vivo". Acta Univ. Palacki Olomouc. Fac. Med. 144, 2000, 82-84.
11. Hampl, R., Lapčík, O., HillI, M., Klak, J., Kasal, A., Nováček, A., Šterzl, I., Šterzl, J.,
Stárka L. 7b-7 Hydroxydehydroepiandrosteron - a Natural Antiglucocorticoid and a
Candidate for Steroid Replacement Therapy? Physiological Research, Suppl 1, 2000,
107-112.
12. Sánchez, D., Tučková, L., Šebo, P., Michalak, P., Whelan, A., Šterzl, I., Jelínková, L.,
Havrdová, E., Beneš, Z., Krupičková, S.,Tlaskalová-Hogenová, H. Increase of IgA
and IgG autoantibodies to calreticulin in coeliac disease: a comparison with various
autoimmune diseases. J. Autoimmunity, 15, 2000, 441-449.
13. Šterzl, I., Hampl, R., Šterzl, J., Votruba, J., Stárka, L. 7b-OH-DHEA counteracts
dexamethasone induced suppresion of primary immune response in murine
splenocytes. Journal of Steroid Biochemistry and Molecular Biology, 1999, 71, str.
133 - 137
14. Šterzl, I., Procházková , J., Hrdá, P., Bártová, J., Matucha, P., Stejskal, V. Mercury
and nickel allergy: risk factors in fatigue and autoimmunity. Neuroendocrinology
Letters, 1999, 20, 221-228.
15. Šterzl, I., Hrdá, P., Procházková, J., Bártová, J., Matucha, P. Reakce na kovy u
pacientů s chronickou únavou a autoimunitními endokrinopatiemi. Vnitřní lékařství
1999,45,č.9,527-531
16. Šterzl, I., Milerová, J., Votruba, J., Šterzl, J. Effect of protein kinase inhibitors on
primary antibody induction in tissue cultures. International Journal of
52
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Immunopharmacolocgy 1998, 20, 583-587
17. Procházková,J., Ivašková,E.,Bártová, J., Šterzl, I., Stejskal,V.Immunogenetic findings
in patients with altered tolerance to heavy metals. Eur.J.Hum.Gnet. 1998 ,6,Suppl 1,
P6.001, 175
18. Šterzl, I., Milerová, J., Votruba, J., Šterzl, J. Effect of protein kinase inhibitors on
primary antibody induction in tissue cultures. The Immunologist. 1998, supp.1, str.
410
19. Šterzl, I., Fučíková, T., Hrdá, P., Matucha, P., Zamrazil, V. Únavový syndrom u
autoimunitní tyreoiditidy s polyglandulární aktivací autoimunity. Vnitřní lékařství
1998, 44, č. 8, 456-460
J.Vencovský
Grants:
1996-1998 - Role CD23 receptoru u revmatoidní artritidy a možnosti blokády CD23 a jeho
ligandů pomocí monoklonálních protilátek (IGA MZ ČR - 3644-3).
1996-1998 - Léčba polymyozitidy a dermatomyozitidy methotrexatem a cyklosporinem A a
nové možnosti hodnocení aktivity onemocnění (IGA MZ ČR - 3638-3).
1. Studium klonality a použití genů variabilních oblastí imunoglobulinů B lymfocytů
revmatoidní synovie na úrovni jedné buňky.
2. Charakterizace neznámých antinukleárních protilátek a jejich příslušného autoantigenu
(vedení subprojektu spolu s prof. Raškou).
3. 1999 – 2001. Mechanizmy tkáňového zánětu u polymyozitidy dermatomyozitidy (IGA
MZ ČR – NI/5369-3).
4. 2001 – 2003 Role vybraných chemokinů, chemokinových receptorů a interferonu beta
v patogenezi zánětu u revmatoidní artritidy (IGA MZ ČR – NI/6327-3).
5. 2003 – 2006 PROMETHEUS - Polymyositis and Dermatomyositis Research on
Methotrexate in European Study (EULAR).
Publications:
1. Hajeer AH, Lazarus M, Turner D, Mageed RA, Vencovsky J, Sinnott P, Hutchinson
IV, Ollier WER. IL-10 gene promoter polymorphisms in rheumatoid arthritis. Scand J
Rheumatol 1998; 27:142-5.
2. Vencovský J. Autoimunitní systémová onemocnění – minimum pro praxi. Triton
1998. 144 s.
3. Mageed RA, Vencovský J, Youinou P, Lydyard PM. Polyreactivity and
immunoglobulin variable region gene usage in human CD5+ B cells. Trans-Caucasian
J Immunol 1999; 1:13-30.
4. Vencovský J, Jarošová K, Macháček S, Studýnková J, Kafková J, Bartůňková J,
Němcová D, Charvát F. Cyclosporine A versus Methotrexate in the treatment of
polymyositis and dermatomyositis. Scand J Rheumatol 2000; 29: (2) 95-102.
5. Brouwer R, Hengstman GJD, Vree Egberts W, Ehrfeld H, Bozic B, Ghirrardello A,
Grondal G, Hietarinta M, Isenberg D, Kalden JR, Lundberg I, Moutsopoulos H, RouxLombard P, Vencovsky J, Wikman A, Seelig HP, van Engelen BGM, van Venrooij
WJ. Autoantibody profiles in the sera of European patients with myositis. Ann Rheum
Dis 2001;60:116-123.
6. Vencovský J, Jarošová K, Růžičková Š, Němcová D, Niederlová J, Ozen S,
53
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Alikasifoglu M, Bakkaloglu A, Ollier WE, Mageed RA. Allele 2 of interleukin-1
receptor antagonist gene is more frequent in patients with juvenile idiopathic arthritis.
Arthritis Rheum 2001;44:2387-2391.
7. Vencovský J, Žďárský E, Moyes SP, Hajeer A, Růžičková Š, Cimburek Z, Ollier WE,
Maini RN, Mageed RA. Polymorphism of the immunoglobulin heavy chain variable
region VH1-69 gene contributes to rheumatoid arthritis susceptibility in HLA-DRB1
shared epitope negative Czech patients. Rheumatology 2002;41:401-410.
8. Ozen S, Alikasifoglu M, Bakkaloglu A, Duzova A, Jarošová K, Němcová D, Besbas
N, Vencovský J, Tuncbilek E. Tumor necrosis factor  G-->A -238 AND G-->A -308
polymorphisms in juvenile idiopathic arthritis. Rheumatology 2002;41:223-227.
9. Vencovský J. Autoprotilátky. In: Tlaskalová-Hogenová H, Holáň V, Bilej M Eds.
Buněčné a molekulární základy imunologie 2003. Česká imunologická společnost,
Praha 2003:141-149.
10. Vencovský J, Šedová L, Macháček S, Kafková J, Gatterová J, Pešáková V, Růžičková
S. Autoantibodies can be prognostic markers of an erosive disease in early rheumatoid
arthritis. Ann Rheum Dis 2003;62:427-430.
11. Isenberg DA, Allen E, Farewell V, Ehrenstein MR, Hanna MG, Lundberg IE, Oddis
C,Pilkington C, Plotz P, Scott D, Vencovsky J, Cooper R, Rider L, Miller F, For
International consensus outcome measures for patients with idiopathic. inflammatory
myopathies. Development and initial validation of myositis aktivity and damage
indices in patients with adult onset disease. Rheumatology (Oxford). 2003 Jul 16
[Epub ahead of print].
P. Kučera
Grants:
1. IGA NI/6349-3. Test aktivace bazofilů a imunoblotting u alergie na jed blanokřídlého
hmyzu 2000 – 2002. Vedoucí řešitel: MUDr. P. Kučera. Hodnocení: B
2.
Specifická imunoterapie - klinické a imunologické aspekty kontinuálně prováděné
léčby 1995 – 1997. Vedoucí řešitěl: MUDr. E. Antošová, spoluřešitel: MUDr. P.
Kučera. Hodnocení: B
Publications:
1. Expoziční test jedem blanokřídlého hmyzu – vlastní zkušenosti. Kučera P, Cvačková M,
Jůzová O, Pachl J. Alergie, 2002, 3, 192-194. ISSN 12123536 IGA NI/6349-3
2. Expoziční test jedem blanokřídlého hmyzu – metodika. Kučera Petr, Cvačková M, Pachl
J, Alergie, 2002, 1,23-25. ISSN 12123536 IGA NI/6349-3
3. Is the basophil stimulation more predictive for bee venom reactivity than antibody
analysis? Kučera P, Cvačková M, Jůzová O, Pachl J. Abstract Book, XXII Congress of
the European Academy of Allergology and Clinical Immunology, Paris, 7-11-June, 2003
4. Peripheral blood basophil activation and immunoblotting in patients with allergy to bee
venom. Kučera P (G), Cvačková M, Pachl J, Juzova O. Poster Abstract XXI Congress of
the European Academy of Allergology and Clinical Immunology, 1-5 June, 2002, Naples.
Allergy, 2002, 57, Suppl. 73, 123, ISSN 0105-4538, Abstrakt
5. HLA in Czech adult patients with autoimmune diabets mellitus:comparison with Czech
children with type 1 diabetes and patients with type 2 diabetes. Cerna M, Novota
P.Kolostova K,Cejkova P, Zdarsky E, Novakova D,Kucera P,Novak J, Andel M. Eur J
Immunogenet, 2003, 30, 6, 401-7
54
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6. Expoziční test jedem blanokřídlého hmyzu. Kučera P (G), Cvačková M, Jůzová O, Pachl
J. Abstrakta 19. Sjezdu ĆSAKI, 6.-9.10.2002, Alergie, 2002, 4, Suppl. 3, 35
7. Pohunek P., Kučera P, B. Suková, F. Votava, J. Zikán Serum ECP taken in the acute
episode of bronchial obstruction can predict the development of bronchial asthma in
young children. Allergy and Asthma Proceedings, 2001, 22, 2, 75-79
8. Kučera P. Nové směry v imunoterapii alergických chorob. Alergie, 2001, 2, 138 – 143
9. Kučera P., M. Cvačková, Test stimulace bazofilů, Alergie,1999, 2, 67-70
10. P. Kučera, E. Antošová, Alternativní schémata imunoterapie alergenem, Alergie, 1999, 3,
159-161
11. P. Kučera, Vernerová E, Antošová E., Specifická imunoterapie - Klinické a imunologické
aspekty kontinuálně prováděné léčby. Klinická imunológia a alergológia, 1998, 1, 9 – 14
P. Panzer
Grants:
1.
Specifické IgE - srovnání technik stanovení. IGA MZ ČR 4717/2. V. Krauz, P.
Panzner, M. Haschová
2.
Sledování specifických protilátek v průběhu alergenové imunoterapie u
pacientů s přecitlivělostí na jed blanokřídlého hmyzu. IGA MZ ČR NI 6825/3. M.
Haschová, P. Panzner, J. Hanzlíková, I. Malkusová
3.
Sledování buněčných a humorálních markerů zánětu v indukovaném sputu
pacientů s alergickým astmatem a alergickou rýmou. IGA MZ ČR NI 7535-3. I.
Malkusová, P. Panzner, J. Hanzlíková, M. Haschová
4.
Je u atopiček menší riziko poruch plodnosti? VZ LF UK 111 4 0000 5. Z.
Ulčová- Gallová, J. Hanzlíková, P. Panzner, I. Malkusová, M. Haschová, M. Liška
Publications
1. Klener P. a kolektiv: Vnitřní lékařství. Alergická onemocnění, p. 426-435.Galén,
Praha, Karolinum, Praha, 1999, 2001.
2. Janů L a kolektiv: Chronický únavový syndrom z pohledu imunologa, internisty,
psychologa a psychiatra. Triton, Praha, 2003.
3. Horký K. a kolektiv: Lékařské repetitorium. Galén, Praha, 2003.
1.
2.
3.
4.
5.
6.
7.
8.
9.
Panzner P.: Možnosti a úskalí diagnostiky alergie s přihlédnutím k imunoterapii.
Alergie, 1, 1999, No. 2, p. 91-94.
Malkusová I., Panzner P., Gutová V., Hanzlíková J., Haschová M.: Švábi - opomíjení
původci alergií a astmatu. Alergie, 3, 2001, No. 3, p. 186-190.
Panzner P.: Cytokiny u chronické obstrukční plicní nemoci a u prosté chronické
bronchitidy. Alergie, 3, 2001, No. 4, 286-295.
Panzner P.: Alergenová imunoterapie. Vox pediatriae, 1, 2001, No. 3, p. 15-18.
Pavelková – Seifertová P., Ulčová – Gallová Z., Balvín M., Netrvalová L., Panzner
P., Rokyta Z.: Význam seminální fruktózy při snížené plodnosti u mužů, vliv na
fertilizaci. Gynekolog, 2001, 10, No. 1, p. 7-9.
Racek J., Holeček V., Sedláček D., Panzner P.: Volné radikály v imunologii a u
infekčních chorob. Epidemiol. Mikrobiol. Imunol., 50, 2001, No. 2, p. 87-91.
Panzner P.: Léčba atopických onemocnění imunoterapií alergenem. Zdravotnické
noviny - příloha Lékařské listy 51, 2002, No. 16, p. 11-14.
Panzner P.: Cytokiny w przewleklej obturacyjnej chorobir pluc oraz przewleklym
zapaleniu oskrzeli. Alergia Astma Imunologia, 8, 2002, No. 2, p. 91-99.
Panzner P.: Diagnostické a terapeutické alergeny. Pomocník alergologa a klinického
imunologa, 2003, p. 176-177.
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10.
11.
12.
Panzner P.:Imunoterapie u alergie a astmatu. Interní medicína pro praxi, 5, 2003, No.
3, p. 131-137.
Panzner P.: Imunoterapie u alergie a astmatu. Interní medicína pro praktické lékaře,
Solen, 2003, p. 33-38.
Panzner P., Lafitte J. J., Tsicopoulos A., Hamid Q., Tulic M. K.: Marked Upregulation of T Lymphocytes and Expression of Interleukin-9 in Bronchial Biopsies
From Patients With Chronic Bronchitis With Obstruction. Chest, 124, 2003, No. 5,
p.1909-1915
R. Špíšek
Publications:
1. Spisek, R., Bougras, G., Ebstein, F., Masse, D., Meflah, K., McIlroy, D. and Gregoire,
M. (2003) Transient exposure of dendritic cells to maturation stimuli is sufficient to
induce complete phenotypic maturation while preserving their capacity to respond to
subsequent restimulation. Cancer Immunol Immunother 52, 445-54. IF 2.8
2. Spisek, R., Chevallier, P., Morineau, N., Milpied, N., Avet-Loiseau, H., Harousseau,
J.L., Meflah, K. and Gregoire, M. (2002) Induction of leukemia-specific cytotoxic
response by cross-presentation of late-apoptotic leukemic blasts by autologous
dendritic cells of nonleukemic origin. Cancer Res 62, 2861-8. IF 8.3
3. Favre, D., Blouin, V., Provost, N., Spisek, R., Porrot, F., Bohl, D., Marme, F., Cherel,
Y., Salvetti, A., Hurtrel, B., Heard, J.M., Riviere, Y. and Moullier, P. (2002) Lack of
an immune response against the tetracycline-dependent transactivator correlates with
long-term doxycycline-regulated transgene expression in nonhuman primates after
intramuscular injection of recombinant adeno-associated virus. J Virol 76, 11605-11.
IF 5.2
4. Pospisilova, D., Borovickova, J., Polouckova, A., Spisek, R., Sediva, A., Hrusak, O.,
Stary, J. and Bartunkova, J. (2002) Generation of functional dendritic cells for
potential use in the treatment of acute lymphoblastic leukemia. Cancer Immunol
Immunother 51, 72-8. IF 2.8
5. Spisek, R., Bretaudeau, L., Barbieux, I., Meflah, K. and Gregoire, M. (2001)
Standardized generation of fully mature p70 IL-12 secreting monocyte-derived
dendritic cells for clinical use. Cancer Immunol Immunother 50, 417-27. IF 2.8
6. Gregoire, M., Ligeza-Poisson, C., Juge-Morineau, N. and Spisek, R. (2003) Anticancer therapy using dendritic cells and apoptotic tumour cells: pre-clinical data in
human mesothelioma and acute myeloid leukaemia. Vaccine 21, 791-4. IF 2.8
7. Spisek, R., Brazova, J., Rozkova, D., Zapletalova, K., Sediva, A., Bartunkova, J.
Bacterial immunomodulators: Clinical-grade maturation factors for the production of
dendritic cell-based vaccines. Vaccine. Přijat k publikaci.
8. Špíšek, R., Bartůňková, J. (2003) Dendritická buňka v imunitě. Vesmír 82, 212-214.
9. Špíšek, R., Bartůňková, J. (2003) Dendritická buňka v protinádorové imunitě. Vesmír
82, 254-256.
10. Pospíšilová, D., Zavacká, A., Hrušák, O., Šedivá, A., Špíšek, R. and Bartůňková, J.
(2000). Dendritické buňky v imunoterapii nádorových onemocnění. Cas Lek Cesk
139, 519-23.
11. Špíšek, R., Bretaudeau, L., Hrušák, O., Poloučková, A., Pospíšilová, D., Bartůňková,
J. (2001) Nové přístupy v imunoterapii-nástup dendritických buněk. Bulletin HPB
56
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Supplement 1, 31-35.
D. Pícha
Publications:
1. Pícha D., Hulínská D., Skalská 2., Moravcová L., Hančil J., Kořínková M., Roháčová
H., Havlíčková J., Hobstová J., Patakiová E., Příhodová J.: Diagnostika lymeské
boréliózy se zaměřením na polymerázovou řetězovou reakci v podmínkách infekční
kliniky. KMIL: 4/1998/6/186-190.
2. Lásiková Š., Pícha D.,: Dynamika lymfocytárních subpopulací v průběhu a po léčbě
lymeské neuroboréliózy. KMIL: 4/1998/6/190-193
3. Lásiková Š., Pícha D.,: Lidská monocytární a granulocytární ehrlichióza. KMIL:
5/1999/1/2-4.
4. Pícha D.: Posudková problematika u méně běžných následků neuroinfekcí. Čas. lék.
českých: 138;1999;19;602
5. Pícha D., Moravcová L., Skalská S., Lásiková Š., Hančil J., Roháčová H., Patakiová
E.: Klinický význam průkazu specifických antiboréliových protilátek v cirkulujících
imunokomplexech u pacientů s lymeskou boréliózou. Klin. Mikrobiol. Inf. Lékařství
6;2000;108-112
6. Lásiková Š., Pícha D., Moravcová L.: Sérové HGE (human granulocytic ehrlichiosis)
protilátky u pacientů s lymekou boréliózou v České republice. . Klin. Mikrobiol. Inf.
Lékařství 6;2000;112-116
7. Žďárský E., Moravcová L., Pícha D.: Detection of Borrelia burgdorferi specific DNA
in urine of patients with Lyme borreliosis. Odesláno k posouzení.
8. Moravcová L, Lásiková Š, Pícha D, Žďárský E. Průkaz specifické DNA Borélia
burgdorferi v moči pacientů s lymeskou boréliózou. Klin.Mikrobiol.Inf.Lék.
2000;7:221-224.
9. Pícha D, Moravcová L, Marešová V. Intrathékální syntéza specifických
antiboréliových protilátek v mozkomíšním moku u pacientů a neuroboréliózou. Čes. a
Slov. Neurol. Neurochir. 2000;63/96:279-282.
10. Moravcová L, Pícha D,Lásiková Š,Marešová V. Stanovení specifických
antiboréliových protilátek v mozkomíšním moku využitím dvou testů obsahujících
odlišné antigeny – Borrelia garinii a Borrelia afzelii. Klin. Mikrobiol.Inf.Lék.:
2001;7:77-79
11. Lásiková Š, Moravcová L, Pícha D, Hančil J,Žďárský E. Detekce specifické boréliové
DNA polymerázovou řetězovou reakcí (PCR) v moči pacientů s lymeskou boréliózou.
Klin.Mikrobiol.Inf.Lék. 2001;7:72-76.
12. Moravcová L, Pícha D, Štěpánová G, Marešová V. Průkaz boréliových antigenů
(vnějšího membránového proteinu OspB a flagelárního proteinu) v mozkomíšním
moku u pacientů s neuroboréliózou. Čes. a Slov. Neurol. Neurochir. 2001;64/97:162167
13. Moravcová L., Lásiková Š., Pícha D.,Žďárský E.: Diagnosticý význam polymerázové
řetězové reakce (PCR) na průkaz Borrelia burgdorferi sensu lato n moči, plazmě a
mozkomíšním moku u pacientů s neuroboréliózou. Klin. Mikrobiol. Inf. Lék.
2002;8;11-14.
Grants:
57
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1) IGA 1761
Lymská borelióza – imunopatogeneze nervových projevů, závislost na věku a význam pro
terapii.
Hlavní řešitel: Dr. Stanislav Doutlík
Spoluřešitelé: Dr. D. Pícha, dr. L. Moravcová, dr. J. Heřmanová, dr. R. Vaněčková, dr. J.
Hančil, dr. H. Roháčová, dr. K. Čech
1993 – 94; hodnocení B
2) IGA 2860
Lymeská neuroborélióza - studium buněčné a humorální imunity a její vztah k průběhu
onemocnění a terapii.
Hlavní řešitel: Dr. Dušan Pícha
Spoluřešitelé: Dr. L. Moravcová, dr. Š. Lásiková
1995 – 97; hodnocení B
3) IGA 4557
Klinický význam průkazu specifických protilátek vázaných v cirkulujících imunokomplexech
izolovaných HPLC u lymeské boréliózy.
Hlavní řešitel: Dr. Dušan Pícha
Spolupřešitelé: Dr. L. Moravcová, dr. S. Skalská, dr. V.Marešová, dr H.Roháčová
1997 – 98; hodnocení B
4) IGA 6244
Diagnostický význam PCR u lymské boréliózy – metodická a klinická studie.
Hlavní řešitel: Dr. Dušan Pícha
Spoluřešitelé: Dr. E. Žďárský, dr. L. Moravcová, dr. S. Skalská, dr. H.Roháčová
2000 – 2; hodnocení
5) IGA 5673
Časná diagnostika neurologických komplikací HIV.
Spoluřešitel
1998-2001
6) Výzkumný záměr 2.LF UK 1113 00003
Komplexní longitudinální klinická a genetická péče o prenatální a postnatální vývoj jedinců.
Koordinátoři: Prof. Goetz, prof. Vavřinec
Hlavní řešitel podúkolu: Role pozitivity průkazu nukleové kyseliny Borrelia b.s.l. v tělních
tekutinách …
1998-2003
Z. Ulčová-Gallová
Publications - Ulčová-Gallová:
Křižanovská K., Ulčová-Gallová Z., Bouše V., Švábek L.,Anděl L.: Vliv některých vlastností
folikulární tekutiny na kvalitu a počet oocytů získaných v programu IVF. Cs.Gynek. 65, 3,
2000, s. 134-138
Ulčová-Gallová Z., Krauz V., Rokyta Z.: Six Kinds of Anti-phopspholipid Antibodies /aPLs/
in Ovulatory Mucus and Seminal Plasma from Couples with Repeated Miscarriages.
Int.J.Fertil. and Wom Dis 45, 4, 2000, p.292-296
Seifertová P., Ulčová-Gallová Z., Panzer P., Křižanovská K., Rokyta Z.: Vyu žití
kryokonzervovaných spermií v mikroaglutinačním estu. Cs.Gynek. 65, 3, 2000, s.138-141
Suchá R., Ulčová-Gallová Z.: Některé vlastnosti folikulární tekutiny. Cs.Gynek.65, 3, 2000, s.
181-188
1. Ulčová-Gallová Z., Mardešic T., Martinez P., Křižanovská K., Rokyta Z.: Some
58
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immunological factors in endometriosis in IVF. Abstr. Book of 7th Biennial World
Congress, London, 14-17 May 2000, N197.
2. Ulčová-Gallová Z.:Některé poruchy plodnosti a protilátky. Zpravodaj Levret klubu, 5,
květen, 2000, s.29-33 ISSN 1212-3846
3. Ulčová-Gallová Z., Bouše V., Rokyta Z., Křižanovská K.: Effect of corticosteroids on
sperm antibody concentration in different biological fluids and on pregnancy outcome
in immunologic infertility. Zentralbl Gynecoll 122, 2000, p. 1-5
4. Ulčová-Gallová Z.: Imunologie v reprodukční medicíně.Postgraduální medicína, 2, 4,
2000, s.466-469 ISSN 1212-4184
5. Iborra A., Palacio J.R., Ulčová-Gallová Z., Martinez P.: Autoimmune Response in
Women with Endometriosis. Amer.J.reprod.Immunol. 2000, p.111-116
6. Pavelková-Seifertová, P.,Ulčová-Gallová Z., Balvín M., Netrvalová L., Panzner P.,
Rokyta Z.: Význam seminální fruktózy při snížené plodnosti u mužů, vliv na
fertilizaci. Gynekolog, 2001, 10, 1, s.7-9
7. Ulčová-Gallová Z.: Komentář k článku Imunizace v těhotenství, Gynekologie po
promoci, I/4, 2001, s. 35, ISSN: 1213-2578
8. Ulčová-Gallová Z., Bouše V., Křižanovská k., Balvín M., Rokyta Z., Netrvalová L.:
Beta2-Glycoprotein I Is a Good Indicator of Certain Adverse Pregnancy Conditions.
Int.J.Fertil and Women Dis, 2001 46, (6), s.304-308
9. Ulčová-Gallová Z.:Poruchy plodnosti očima reprodukčního imunologa. Forum
medicinae, od lékařské vědy k praxi, 2001, 5-6, s.105-109
10. Ulčová-Gallová Z.: Čekání na Kateřinu, Praha, Petrklíč , 2001, s.1-120
11. Ulčová-Gallová Z., Bouše V., Švábek L., Turek J., Rokyta Z.: Endometriosis in
reproductive Immunology. AmJ.Reprod.Immunol. 2002, 47, s.269-274
12. Vančíková Z., Chlumský V., Sokol D., Horáková D., Hamšíková E., Fučíková T.,
Janatková I., Ulčová-Gallová Z., Štěpán J., Limanová Z., Kocna P., Sanchez D.,
Tučková L., Tlaskalová-Hogenová H.: The prevalence of seropositivity for celiac
disease in the general population and in some at risk groups of adults in the Czech
Republic-preliminary report. Folia Biol. 47,6, p.753-758
13. Šemberová J., Pískatá M., Ulčová-Gallová Z., Křižanovská K., Netrvalová L.,
Kučerová L., Rokyta Z.: Penetrace nativních a kryokonzervovaných spermií
ovulačním hlenem-Kremerův test. Čs.Gynekologie 67, 2002, 2, s.89-92
14. Suchá R., Ulčová-Gallová Z., Pavelková-Seifertova P., Křižanovská K., Bouše V.,
Švábek L., Rokyta P., Balvín M., Pecen L., Rokyta Z.: Fruktóza a glukóza ve
folikulární tekutině a v séru stimulovaných žen v programu IVF. Cs.Gynek. 67, 2002,
3, s.144-148
15. Suchá R., Ulčová-Gallová Z., Pikner R., Bouše V., Křižanovksá K., Topolčan O.,
Švábek L., Rokyta P., Rokyta Z.: Dlouhý a krátký stimulační protokol v IVF:
hormonální hladiny ve folikulární tekutině a v séru. Praktická Gynek. 4/2002, s.22-25
16. Ulčová-Gallová Z.: Imunologická příčina poruchy plodnosti. Moderní Gynek. a
porodnictví 11, 2002, 4, s.553-560
17. Ulčová-Gallová Z.:Annexin V a protilátky proti annexinu V v souvislosti se
selháváním lidské reprodukce. Moderni Gynek. a porodnictví 11, 2002, 4, s.561-564
18. Křižanovská K., Ulčová-Gallová Z., Bouše V., Rokyta Z.: Hmotnost a poruchy
reprodukce. Sborník Lék., vol, 103, 4, 2002, s. 517-526
19. Přibylová L., Podzimek Š.,Ulčová-Gallová Z., Procházková J., Bártová J., Rokyta Z.:
Protilátky proti spermiím a imunologická intolerance některých kovů u neplodných
párů. Čs.Gynek. 68, 2003, 2, s.106-110
20. Podzimek A., Procházková J., Přibylová L., Bártová J., Ulčová-Gallová Z., Mrklas L.,
Stejskal V.D.M.: Vliv těžkých kovů na imunitní reakci u pacientů s prokázanou
59
RP identification code
MSM0021620812
neplodností. Čas.Lék.čes. 142, 2003, s.285-288
21. Ulčová-Gallová, Z.: Poruchy plodnosti z pohledu reprodukčního imunologa a
gynekologa.Alergie, roč.5, 2, 2003, s.42-46
22. Beranová m., Šíma P., Rokyta Z., Ulčová-Gallová Z., Vaněček T., Šíma R.:On the
search for the genetic background of thr primary idiopathic infertility:the leukemiainhibitory factor gene mutations in the population of infertile women. Fertil. Steril.
2003, 80 (S3): 89-90
23. Z.Shoenfeld, Krause I., Kvapil F., Sulkes J., Lev S., P.von Landenberg, J.Font,
J.Zaech, R.Cervera, J.C.Piette, M.C.Boffa, M.A.Khamashta, M.L.Bertolaccini,
G.R.Hughes, P.Youinou, P.L.Meroni, V.Pengo, J.D.Alves, A.Tincani, G.Szegedi,
G.Lakos, G.Sturfelt, A.Jonsen, T.Koike, N.Sanmarco, A.Ruffatti, Z.Ulcova-Gallova,
S.Prapornik, B.Rozman, M.Lorber, V.B.Vriezman, M.Blank.: Prevalence and Clinical
Correlations of Antibodies against Six Beta2-Glycoprotein I-related peptides in the
antiphospholipid syndrome. J.Clinic.Immunology, 23, 5, 2003, s. 377-384
24. Semberova J., Ulčová-Gallová Z., Manthay A., Pískata M., Balvín M., Milichovksá
L., Bouše V., Krizanovská K., Rokyta Z.: ELISA Detection of Antichlamydial
Antibodies in Non standard Biological Fluids and their Relationship to female
Infertility. Clinical Application of Immunol., 2, 2003, s. 250-254
P. Dřevínek
Publications:
1. P. Dřevínek, O. Cinek, J. Melter, L. Lanšádl, Y. Návesňáková, V. Vávrová:
Genomovar distribution of Burkholderia cepacia complex significantly differs
between Czech and Slovak patients with cystic fibrosis. J Med Microbiol, 2003, 52(7):
603-604.
2. P. Dřevínek, H. Hrbáčková, O. Cinek, J. Bartošová, O. Nyč, A. Nemec, P. Pohunek:
Direct PCR detection of Burkholderia cepacia complex and identification of its
genomovars by using sputum as source of DNA. J Clin Microbiol, 2002, 40(9): 34853488.
3. Z. Sumnik, P. Drevinek, V. Lanska, H. Malcova, J. Vavrinec, O. Cinek: Higher
maternal age at delivery, and lower birth orders are associated with increased risk of
childhood type 1 diabetes mellitus. Exp Clin Endocrinol Diabetes, 2003, in press.
4. Z. Šumník, P. Dřevínek, M. Šnajderová, S. Koloušková, P. Sedláková, M. Pechová,
J.Vavřinec, O. Cinek: HLA-DQ polymorphisms modify the risk of thyroid
autoimmunity in children with Type 1 diabetes. J Pediatr Endocrinol Metab, 2003,
16(6):851-8.
5. O. Cinek, P. Dřevínek, Z. Šumník, B. Bendlová, P. Sedláková, S. Koloušková, M.
Šnajderová, J. Vavřinec: The NeuroD polymorphism Ala45Thr is associated with
Type 1 diabetes mellitus in the Czech population. Diabetes Res Clin Pract, 2003,
60(1): 49-56.
6. O. Cinek, P. Dřevínek, Z. Šumník, B. Bendlová, S. Koloušková, M. Šnajderová, J.
Vavřinec: The CTLA4 +49 A/G dimorphism is not associated with Type 1 diabetes in
Czech children. Eur J Immunogenet, 2002, Jun; 29(3): 219-22.
7. P. Dřevínek, Z. Šumník, O. Cinek: [Environmental factors in ethiology of childhood
type 1 diabetes mellitus] (Review, in Czech). Diabetes, metabolismus, endokrinologie
a vyziva (Prague), 2001, 4 (4), 270-276.
H. Marečková
Grants:
1. IGA MZd ČR 3120-3 Imunologické abnormality u pacientů s chronickým únavovým
60
RP identification code
MSM0021620812
syndromem 1995-1998
2. IGA Mzd 5740-3 Přínos vyšetření produkce intracelulárních cytokinů pro klinickou
imunologiii 1999-2001
Publications:
1. Macurová,H.,Kamínkova,J.,Fučíková,T.,Janatová,I.,Marečková,H.,Poch,T.
Chemiluminiscence test in the laboratory valuation of the immunomodulatice
treatment Časopis Lékařů Českých. 2000; 139(9): 277-279.
2. Kozák,T., Šavrdová,E., Pit'ha,J., Gregora,E., Pytlik,R., Maaloufová,J,, Marečková,H.,
Kobylka,P., Vodvárková,S. High-dose immunosuppressive therapy with PBPC
support in the treatment of poor risk multiple sclerosis. Bone-Marrow-Transplantation.
2000; 25(5): 525-531.
3. Amaraa R., Marečková H., Urbánek P., Fučíková T.: T helper, cytotoxic T
lymphocyte, NK cell and NK-T cell subpopulations in patients with chronic hepatitis
C, 2002,
4. Folia Microbiol, 47 (6), 717-723
5. Amaraa R.,Marečková H.,Urbánek P.,Fučíková T.: Productionof interleukins 10 and
12 by activated peripheral blood monocytes/macrophages in patients suffering from
chronic hepatitis C virus infection with respect to the response to interferon nad
ribavirin treatment.2002, Immunology Letters, 83,(3) 209-214
6. Amaraa R., Marečková H., Urbánek P., Fučíková T.: Zvýšení počtu CD4+
pomocných T lymfocytů v periferní krvi a jejich kompartmentalizace v jaterní tkáni u
pacientů s chronickou hepatitidou C 2002 Česká a slovenská gastroenterologie, 56,3,
88-94
7. Marečková H., Ravdan A.,Fučíková T., Janatková I.: Vyšetření produkce
intracelulárních cytokinů cytokinů T lymfycyty pomocí průtokové cytometrie –
metodické problémy, 2002,Epidemiol Mikrobiol Imunol., 51,3,111-118
8. Marečková H., Fučíková T.:Příznivý efekt Trypsinu retard u nemocné s
leukocytoklastickou vaskulitidou,2002,Praktický lékař, 82, 10,603-605
9. Amaraa,R; Marečková,H., Urbánek,P., Fučíková,T. Immunological predictors of
different responses to combination therapy with interferon < alpha > and ribavirin in
patients with chronic hepatitis C. Journal-of-Gastroenterology. 2003; 38(3): 254-259.
P. Hrdá
Grants:
1. NB/6317-3 Genetické vazby u polyglandulárních autoimunitních endokrinopatií. IGA
MZ ČR, 2000-2002
Publications:
1.
P. Hrdá, I.Šterzl, P.Matucha. Cytokine levels in sera of patients with
autoimmune endocrinopathies. Physiological Research 52, 2003, 256-267.
2.
Šterzl, I., Hampl, R., Hill, M., Hrdá, P., Matucha, P. Immunomodulatory
cytokines in human seminal plasma correlate with immunomodulatory steroids.
Steroids 68(9), 2003, 725-731.
3.
I.Šterzl, P.Hrdá, P.Matucha, F.Korioth, A.Kromminga. Porovnání
autoimunitních endokrinopatií v oblasti exprese HLA antigenů. Alergie 4, 2002, 281284
4.
P.Hrdá, F.Korioth, P.Matucha, I.Šterzl, A.Kromminga. HLA association with
61
RP identification code
MSM0021620812
5.
6.
7.
8.
autoimmune endocrinopathies. In. From Proteomics to Molecular Epidemiology:
relevance of Autoantibodies (Report on the 6 th Dresden Symposium on
Autoantibodies held in Dresden on September 4-7.2002). K. Conrad, M. Fritzler, M.
Meurer, U. Sack, Y. Shoenfeld (eds.), Pabst Science Publishers, Lengerich, Berlin,
Riga, Rom, Viernheim, Wien, Zagreb, 2002, 248-249.
P. Hrdá, I.Šterzl. Autoimunitní polyglandulární syndromy. Alergie, vo.3, 2001,
126-129.
I.Šterzl, P.Hrdá, J.Procházková, J.Bártová, P.Matucha. Reakce na kovy u
pacientů s chronickou únavou a autoimunitními endokrinopatiemi. Vnitřní lékařství
1999,45,č.9,527-531
I.Šterzl, J.Procházková, P.Hrdá, J.Bártová, P.Matucha, V.Stejskal. Mercury
and nickel allergy: risk factors in fatigue and autoimmunity. Neuroendocrinology
Letters, 1999, 20, 221-228.
I.Šterzl, T.Fučíková, P.Hrdá, P.Matucha, V.Zamrazil. Únavový syndrom u
autoimunitní tyreoiditidy s polyglandulární aktivací autoimunity. Vnitř. lékař. 1998 ,
44, č.8, 456-460.
D. Sedláček
1. Sedláček D. Postexpoziční profylaxe infekce HIV. Klin mikrobiol inf lék. 2000;
6(4): 102-105
2. Sedláček D, Stehlík P, Stožický F. Příspěvek ke kontrole adherence léčby
antiretrovirovými přípravky. Klin Mikrobiol Inf Lék. 2001;7(3):80-82
3. Sedláček D, Ulčová-Gallová Z, Milichovská L, Nováková P, Rokyta Z. Seven
Antiphospholipid Aantibodies in HIV-Positive Patiens: Correlation with Clinical
Course and Laboratory Findings. Am. J Reprod. Immunol. 2003. V tisku.
A. Janda
1. A.Šedivá, J.Bartůňková, R.Zachová, O.Hrušák, A.Janda, E.Kočárek, D.Novotná,
K.Novotná, V.Krutílková, T.Klein: Early Development of Immunity in DiGeorge
Syndrome. In: Alergologijos žinijos,3, ISSN 1684-4266. EAACI Summer School,
Vilnius, Litva, 2003.
62
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E
Description of spatial, material and technical guarantee of the
research plan solution
E1.
Assurance of the realisation of the research plan in the framework of existing
spatial, material and technical conditions in the workplace of the
applicant/administrator
In the medical schools the laboratories of high quality were built in the previous years. It
enables them to conduct valuable research in the field of immunology. The planned research
could be accomplished with the use of current equipment. However, the uprade and
improvement of this equipment seems to be necessary.
E2.
Infrastructure, instrumentation and technical equipment which is necessary to
procure in order to realise research plan objectives
The existing devices bought couple years ago are in a way out dated and it is necessary to
upgrade them or even buy new, more productive ones.
63
RP identification code
MSM0021620812
F
F1.
Financial assurance of the research plan solution
Allowable costs in thousands of CZK
Year 2005
Total
Wages and salaries
Agreements to work outside the scope of employment
Compulsory statutory taxes
Allocation into the Fund of Social and Cultural Needs
Property acquisition costs
Costs of depreciation, maintenance and repairs of the
property
Purchase of material, small inventory and stocks
Purchase of services
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
B1
B3
B7
B9
B11
B13
8644
B15
B17
B19
B21
B23
B25
E15
2878
265
470
40
10
737
20858
B16
B18
B20
B22
B24
B26
E16
11701
8656
4101
485
40
10
927
25920
Out of which
institutional support
E20
10079
E22
6564
E24
4101
E26
485
E28
40
E30
10
E32
185
E34
21464
11701
13382
4636
485
30
20
1034
31288
Out of which
institutional support
E38
10079
E40
11290
E42
4636
E44
485
E46
30
E48
20
E50
207
E52
26747
11701
6092
4301
490
30
20
968
23602
Out of which
institutional support
E56
10079
E58
4000
E60
4301
E62
490
E64
30
E66
20
E68
194
E70
19114
Year 2006
3026
31
2285
2472
Total
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
E19
E21
E23
E25
E27
E29
E31
E33
Year 2007
Total
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
E37
E39
E41
E43
E45
E47
E49
E51
Year 2008
Total
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
64
Out of which
institutional support
B2
7443
B4
B8
2605
B10
31
B12
2285
B14
0
E55
E57
E59
E61
E63
E65
E67
E69
2878
265
470
40
10
589
16616
RP identification code
MSM0021620812
Year 2009
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
E73
E75
E77
E79
E81
E83
E85
E87
Year 2010
11707
4092
4000
500
30
20
910
21253
Out of which
institutional support
E92
10079
E94
2000
E96
4000
E98
500
E100
30
E102
20
E104
182
E106
16811
11707
4092
4000
500
30
20
910
21253
Out of which
institutional support
E110
10079
E112
2000
E114
4000
E116
500
E118
30
E120
20
E122
182
E124
16811
Total
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
E91
E93
E95
E97
E99
E101
E103
E105
Year 2011
Total
Personal expenses
Property acquisition costs
Cost of operation
Travel expenses
Costs of international cooperation
Costs of the publication of results and rights to the results
Supplementary (overhead) expenses
Total
F2.
11701
4092
4117
505
30
20
934
21399
Out of which
institutional support
E74
10079
E76
2000
E78
4117
E80
505
E82
30
E84
20
E86
187
E88
16938
Total
E109
E111
E113
E115
E117
E119
E121
E123
Justification of items and allowable expenses, specification of financial resources
65
RP identification code
MSM0021620812
Wages and salaries are calculated according to the regulations of the 2nd Medical School,
Charles Univeristy, and Compulsory statutory taxes account for 35% of the expenses. We
expect that majority of the researchers wil work half-time on behalf of the project, with seven
young ones employed full time. Part of the expenses will be covered from sources of the 2nd
Medical School. The sum in the Material, small inventory and stocks row will be spent to
purchase stationary, reagents, chemicals, and small equipment for the laborator. Services
includes the transportation of samples, statistics, administrative and graphic work. The travel
costs account for the cost of travel in the Czech Republic and abroad. Investment
requirements. during the first years of the research plan, there shall be purchased several
pieces of equipment necessary for the laboratories involved. Cost of depreciation,
maintanance and repairs of the property bought from other sources will be covered by 2nd
Medical School.
Expense calculation strategy for the following years. The total amount of costs varies in
the following years, chiefly deriving from the extent of the investments. It reaches its peak in
the year 2007 when we plan the greatest investment – purchase of laser scan microscop
Compucyte, another great investment is purchase of cytometr FACS in the year 2006. The
level of costs in the following years is lower and stable. In this time we do not plan to buy any
bigger device and the financial sources will be used mainly for maintanance and upgrade of
the existing devices.
66