Download Immunology 03 MED

DEPARTMENT OF MCROBIOLOGY AND IMMUNOLOGY
COURSE IN IMMUNOLOGY FOR STUDENTS OF FACULTY OF MEDICINE
The name of Unit in which the subject is realized
Department of Microbiology and Immunology
Head
Prof. dr hab. Stefania Giedrys-Kalemba
Total hours:
60 hours include:
15 h of seminars
45 h classes
Tutor:
dr n. med. Monika Nowosiad
ECTS: 4
Aims of teaching
1. to become acquainted with a structure and function of the immune system
2. to learn about mechanisms of immune response to different antigens (bacteria, fungi, viruses,
graft, tumor cells, allergens)
3. to recognize and detect of immune response reactions in vitro and in vivo
4. to know how to interpret immunological tests results
5. to get knowledge of immune system modulation methods (vaccination, serotherapy,
nonspecific immunotherapy, desensitization)
FORMS OF ACTIVITIES
Immunology is Vth semester subject, conducted in such forms: classes, seminars.
1.
2.
3.
Seminars are conducted during 15 h. During the seminars the theory is presented and
discussed with using slides and sometimes films.
Practical classes include 45 h. Seminars and classes are lasting together 4 h. During the
classes students are controlled by tutor with acquaintance of subject theory (short tests: 10
questions, 10 min), watch, prepare and analyze different tests etc. The classes are carried out
in 10-11 group of students per tutor. Students are obliged to use only those protective coats,
which are available in our department. Lessons are obligatory. The topics are given prior to
the lessons on the notice board. Every absence has to be justified and the lesson has to be
made up in the microbiological laboratory and passed theoretically.
The final credit is conducted in a written form - one-choice test. All lessons have to be
completed before the final credit.
Program of immunology
1. Introduction to the immune system. Nonspecific immunity I.
The lymphoid system: primary (central) and secondary (peripheral) lymphoid organs. Cells of the immune system and
their functions: stem cell, B, T, NK lymphocytes, macrophages, granulocytes, dendritic cells, mast cells, platelets.
Soluble mediators: complement, antibodies, cytokines, interferons, inflammatory mediators.
Nonspecific immunity: exterior defenses and physical and biochemical barriers, the role of normal flora, nonspecific
factors humoral (complement, interferons, lysozym, lactofferin, C-reactive protein, heat shock proteins..) and cellular
(mononuclear and polymorphonuclear phagocytes, NK cells). Pathological barrier - inflammation.
Complement: classical and alternative pathways, biological effects (increase of vascular permeability, chemotaxis and
neutrophils activation, opsonization, lysis). Complement receptors.
Practically
Film: The immune system.
Measurement of complement activity – detection of individual components: C3, C4, C1 inhibitor, B, P factors and
hemolytic activity: lysis of 50% of a standardized preparation of antibody-sensitized erythrocytes - CH50.
Estimation of chemotaxis – agarose method.
2. Nonspecific immunity II.
Immunity: innate and adaptive (acquired) active and passive, specific and nonspecific, natural and artificial, cellular and
humoral. Immunity and immune response.
Phagocytosis: migration and chemotaxis of phagocytes, adhesins molecules (integrins, selectins), chemotactic factors
(complement proteins, chemokines), phagocytes receptors, opsonization, ingestion, digestion (killing), oxygen-dependent
and oxygen-independent microbicidal activity.
Natural cytotoxicity – NK cells.
Practically
Film: Local pulmonary defense mechanism.
Assays for phagocyte cells – percentage of phagocytes, index of phagocytosis, index of killing, NBT
(nitrobluetetrazolium test).
3. Specific immune response I (cellular).
Antigen, hapten, chemical structure, thymus-dependent and thymus-independent antigens, cross-reactivity antigens.
Antigenic determinants – epitopes, immunogenicity, specificity.
Main phases of the immune response: induction phase (recognition of antigen), central phase
(activation, clonal selection and prolipheration of T and B lymphocytes), effector phase (elimination of antigen mediated
by antibodies and effector cells). Immunological memory and tolerance.
Lymphocytes: subpopulations: B (B1, B2), T (Th1, Th2, Ts, Tc), NK, NC, CD markers, receptors for antigen (B – Ig, T –
TCR), circulation of lymphocytes.
Processing and presentation of antigen, antigen presenting cells.
Specific cellular response: cell-mediated cytotoxicity (recognition of antigen: T CD8 – I class MHC restriction), delayed
type of hypersensitivity (T CD4 – II class MHC restriction, effector phase – activated macrophage). The cytokine
network.
Practically
Film: Cellular mechanisms of the immune response.
Estimation of level and function of T and B lymphocytes: separation of lymphocytes, detection of the CD markers
(rosetting tests: E, EA, EAC, differentiation of lymphocytes with using IF, flow cytometry), lymphocytes functions
testing (activation and prolipheration after PHA, migration inhibitory test, concentration of cytokines, cytotoxic tests).
4. Specific immune response – II (humoral).
Specific humoral response: B lymphocytes recognition, T and B cooperation in the antibody response, plasma cells –
antibody production, primary and secondary humoral response.
Cooperation of specific humoral and cellular response: immunophagocytosis, antibody dependent cellular cytoxicity
(ADCC) – NK CD16, macrophages, neutrophils
Appropriate (defense against infections, pre-cancer growth control) and inappropriate (allergy, autoimmunity, transplant
rejection) specific response consequences.
Antibodies: structure, Fab and Fc role, sequence differences (isotypic, allotypic, idiotypic, paratop), biological effector
functions, Fc receptors on cells, monoclonal and idiotypic antibodies, specificity, affinity, avidity, cross-reaction. Types
of immunoglobulins.
Antigen-antibody interactions: in vivo – neutralization, immunological complexes, opsonization; in vitro – agglutination,
precipitation, passive haemagglutination, complement fixation, direct and direct immunofluorescence, Elisa, RIA,
immunoblotting
Practically
Film: Antibody structure and the generation diversity
Immunoglobulins classes (IgG, IgM, IgA) - estimation in serum with single radial immunodiffusion.
Detection of specific antibodies and antigens in serological tests in vitro: slide and tube agglutination, ring precipitation,
double gel diffusion, radial diffusion, lytic test, complement fixation test, If, Elisa, immunoblotting.
5. Ontogeny and phylogeny of the immune system. Infection immunity.
Filogenesis and ontogenesis of immune system: development and maturation of immune system: fetus, newborn, child,
adult, old age, immunobiology of aging. Influenced factors.
The immune system of skin and mucosa- SALT, MALT- GALT, NALT, BALT -similarity and diversity, food tolerance.
Regulation of the immune response (role of antigen molecules which specifically recognize antigen, immunoglobulins,
the generation of diversity of antibodies, T-cell antigen receptors, idiotypic antibodies). Neuroendocrine modulation.
Immunological tolerance, mechanisms.
Immune response for different antigens. Types of infections and parasites: obligatory intracellular parasites, facultative
intracellular parasites, extracellular parasites.
Immunity to bacteria (immune response to extracellular and intracellular bacteria, bacterial evasion of host-defense
mechanisms), fungi, viruses (viral neutralization by humoral antibody, cell-mediated and humoral antiviral mechanisms),
protozoa and worms. Role of structure, metabolism, division, specific and nonspecific defense. Escape mechanisms.
Practically:
Film: Infectious disease.
Results of immunological examinations of patients of different age –analysis.
The results of serological tests in different infections: Helicobacter pylori, Borrelia burgdorferi, Mycoplasma
pneumoniae, Chlamydia pneumoniae, Ebstein-Bar virus.
6. Hypersensitivity.
Mechanisms of hypersensitivity. Early reactions: type I – anaphylaxis, allergens, IgE antibodies, IgE receptors, involved
cells (mast cells, basophiles), mediators, clinical effects (hay fever, asthma, eczema, anaphylaxis); type II – cytotoxic and
cytolytic reactions (posttransfusion, drug-induced reactions); type III – immune-complex diseases (Arthus reaction,
serum sickness); late reactions: type IV – tuberculin (bacterial allergy, contact hypersensitivity)
Practically
Estimation of IgE in vitro – RIST and RAST tests. Microscopic observation of eosynophils in bronchoalveolar lavage
(BAL) preparations. Direct and indirect Coombs test.
Late hypersensitivity in people – Multitest.
7. Autoimmunity.
Autoimmunity: autoantigens, autoantibodies, factors influence on disorders of self-antigens tolerance. Mechanisms of
autoimmune diseases arising – type II and III hypersensitivity, influence of genetic factors, specificity of autoimmune
response – organ-specific and non organ-specific diseases. Antigenic mimicry (rheumatic fever).
Positive role of IgE, autoimmunity and tolerance.
Practically
Film. Autoimmune disease.
The rules of autoimmune diseases diagnostics - autoantibodies detection- IF method, ELISA, Western -blot and immune
complexes detection.
8. Primary and secondary immunodeficiency
Primary immunodeficiency: B-cell dependent , T-cell dependent, defects in complement proteins and in phagocytes).
Secondary immunedeficiency: caused by drugs, nutrition, other diseases (AIDS).
Infections typical for different types of immunodeficiency.
Practically
Analysis of immunological parameters in health and immunodeficiency.
9. Transplantation and graft rejection
Transplantation immunology: general organization and inheritance of the MHC/HLA complex, transplantation antigens
HLA class I and II, bone marrow transplants, organ transplants, relationship between the donor and recipient,
immunologic mechanisms involved in allograft rejection graft-versus-host response (GVHD).
HLA antigens and susceptibility to diseases.
Practically
Film: Transplantation
HLA antigens class I and II typing: serological methods, molecular methods (PCR-SSP, PCR-SSO). Limphocytotoxic
test (LCT). Donor-recipient maching.
10. Immunoprophylaxis, immunomodulation, immunotherapy.
Active immunization: types of vaccines – whole organisms inactiveted and attenuated, purified molecules (toxoids,
capsular polysaccharide, surface antigens), recombinant antigen vaccines, DNA vaccines; recommended vaccinations,
indications and contraindication and side effects. Vaccinations in groups of risk. Adjuvants - mechanism of action.
Recomenndated vaccines - group of risk
Passive immunization: indications, complications.
Autovaccines – indications, way of administration.
Desensitization: vaccines used in the atopic diseases.
Non-specific vaccines and immunotherapy (cytokines).
Nonspecific immunotherapy (bacterial, plantal, cytokines), immnosupression.
Practically:
Demonstration of different vaccines, sera and immunomodulating preparations.
Autovaccines – indications and contraidications, principles of treatment.
11. Tumor immunology and immunology of reproduction.
Cancer and the immune system: malignant transformation of cell, oncogens and cancer induction, tumor antigens,
differentiation of tumor-antigens, human immune response to tumor and escape mechanisms. Immunotherapy of tumors.
Immunology of reproduction: immunological base of infertility in man and woman, gravidity as an allogenic transplant,
pregnancy maintenance, pregnancy loss, immunotherapy of recurrent abortions.
Practically:
Film: Monoclonal antibodies.
Immunoprophylaxis of Rh (-) women; administration of anti-D immunoglobulin.
Antisperm antibodies in IF method.
Cytotoxity of NK cells (L-1210 leukemia).
LAK cells cytotoxity.
Disscussion of clinical cases of immunological diseases. After transplantation, autoimmune diseases, cytopenia, lupus
eryhematosus, hepatitis, thyreoditis.
12. 13. Immunological diseases diagnostic procedures.
Practically:
Discussion of clinical cases of immunological diseases. After transplantation, autoimmune diseases, cytopenia, lupus
eryhematosus, hepatitis, thyreoditis etc.
14. Made up the classes can for the absence
15. Final credit test.
Recommended textbooks:
Immunology, I. Roitt, J. Brostoff, D. Ihale, Mosby, 7th ed. 2006, ISBN 0323033997
Immunology, R. M. Hyde, 3rd or 4th edition, NMS