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Virchows Arch (2006) 449: 96–103
DOI 10.1007/s00428-006-0176-7
ORIGINA L ARTI CLE
Christina Mayerl . Melanie Lukasser . Roland Sedivy .
Harald Niederegger . Ruediger Seiler . Georg Wick
Atherosclerosis research from past to present—on the track
of two pathologists with opposing views, Carl von Rokitansky
and Rudolf Virchow
Received: 21 November 2005 / Accepted: 9 February 2006 / Published online: 13 April 2006
# Springer-Verlag 2006
Abstract It is now clear that inflammation plays a key role
in atherogenesis. As a matter of fact, signs of inflammation
of atherosclerotic plaques have been observed for centuries
and also constituted the basis for a fierce controversy in the
19th century between the prominent Austrian pathologist
Carl von Rokitansky and his German counterpart, Rudolf
Virchow. While the former attributed a secondary role to
these inflammatory arterial changes, Virchow considered
them to be of primary importance. We had the unique
opportunity to address this controversy by investigating
atherosclerotic specimens from autopsies performed by
Carl von Rokitansky up to 178 years ago. Twelve
atherosclerotic arteries originally collected between the
years 1827 to 1885 were selected from the Collectio
Rokitansky of the Federal Museum of Pathological Anatomy, Vienna Medical University. Using modern sophisticated immunohistochemical and immunofluorescence
techniques, it was shown that various cellular intralesional
components, as well as extracellular matrix proteins, were
preserved in the historic atherosclerotic specimens. Most
importantly, CD3 positive cells were abundant in early
lesions, thus, rather supporting Virchows’s view, that
inflammation is an initiating factor in atherogenesis.
Furthermore, we hope to have opened a new and intriguing
possibility to study various pathological conditions using
valuable historical specimens.
C. Mayerl . M. Lukasser . H. Niederegger . G. Wick (*)
Division of Experimental Pathophysiology and Immunology,
Department Biocenter, Innsbruck Medical University,
Fritz-Pregl Str. 3,
6020 Innsbruck, Austria
e-mail: [email protected]
Tel.: +43-512-5073100
Fax: +43-512-5072867
R. Sedivy
Department of Clinical Pathology, University Hospital Vienna,
Vienna, Austria
R. Seiler
Department of Vascular Surgery,
University Hospital Innsbruck,
Innsbruck, Austria
Keywords Atherosclerosis . Inflammation . Historical
material
Introduction
Atherosclerosis is a disease that has plagued mankind for
millennia, as exemplified by findings of degenerative
changes in the aorta, coronary, and peripheral arteries of
Egyptian mummies [13, 14, 27]. These lesions from
ancient Egyptians are no different from those currently
observed in vascular surgery and morbid histology.
Medical researchers began to take a closer look at
vascular alterations in the beginning of the 19th century. In
1829, the German-born French surgeon and pathologist
Jean Lobstein introduced the term “arteriosclerosis” in his
unfinished Traité d’Anatomie Pathologique, a four-volume
treatise on pathological anatomy, based upon his lifelong
personal experience [6].
In the middle of the 19th century, two opposing schools
of pathology, that of Rudolf Virchow in Berlin, Germany,
and that of Carl von Rokitansky in Vienna, Austria,
described cellular inflammatory changes in atherosclerotic
vessel walls [11, 17]. Although von Rokitansky considered
these changes secondary in nature, Virchow supported their
primary role in atherogenesis. Contrary to the “humoral
pathology” and the “crases” theory of the Viennese school,
Virchow saw the causes of diseases in changes of cells and
coined the term “cellular pathology”.
Virchow strongly criticized Carl von Rokitansky and the
Viennese Medical School for their dogmatism and support
of outdated humoralism. In fact, the humoral disease theory
included in the first edition of Rokitansky’s famous
pathology textbook “A Manual of Pathological Anatomy”
was eliminated entirely from the second edition.
In 1910, the German chemist Windaus showed that
atherosclerotic plaques consist of calcified connective
tissue and cholesterol [23]. Three years later, Anitschkow
and Chaltow succeeded in inducing atherosclerosis in
rabbits by feeding a cholesterol-rich diet [1] unequivocally
identifying a classical risk factor for progression of
97
atherosclerotic alterations. Several other risk factors, such
as smoking, high blood pressure, high emotional stress,
improper diet, diabetes, etc., were later found to contribute
to disease progression.
However, the crucial first pathogenetic events of the
disease remained unclear. Classical concepts of atherogenesis did not attribute a major relevance to inflammatoryimmunologic processes as possible pathogenetic factors. A
comprehensive review on historical papers dealing with
inflammatory aspects of atherosclerosis has been compiled
by Nieto [9].
The “response to injury hypothesis”, summarized by
Ross in 1993, originally postulated an alteration of the
endothelium and intima due to, e.g., mechanical injury,
toxins, and oxygen radicals, as the initiating event leading
to endothelial dysfunction [12].
Alternatively, the “altered lipoprotein hypothesis”,
favored by Steinberg and others, postulated an initiating
role of chemically altered lipoproteins, notably oxidized
low-density lipoprotein (oxLDL), leading to the primary
formation of foam cells in the intima [16]. The group of
Fogelman [8] pointed out that native LDL, rather than
oxLDL, is transported into the intima through the endothelium, where it is then modified and retained, acting as a
chemoattractant for monocytes and smooth muscle cells
(SMC), and being later taken up by these cells, resulting in
foam cell formation. This concept was termed the “retention of modified LDL hypothesis”. However, atherogenesis
seems to be a far more complex process than simple lipid
deposition.
Intriguingly, accumulations of activated T cells, macrophages, and dendritic cells were identified in severely
afflicted arteries [2] and were also found to be common in
arteries of young adults and even children, at predilection
sites for the later occurrence of atherosclerotic lesions [18,
25]. We have called these foci of intimal mononuclear cells
the “vascular-associated lymphoid tissue”—VALT [7, 22].
With respect to these findings, the question arose, as to
which antigens are recognized by the immune system,
leading to a local immune reaction in the arterial wall. An
extended series of experiments revealed a stress protein,
heat shock protein 60 (HSP 60), as the potential (auto)
antigen inciting an immune response at the beginning of
atherosclerotic changes in the vessel wall [4, 24]. Classical
atherosclerosis risk factors, such as high blood pressure,
smoking, diabetes, biochemically modified LDL, etc.,
result in the expression of HSP60 by endothelial cells (EC),
especially at sites that are subjected to major (turbulent)
haemodynamic stress and known to be predilection sites
for the later development of atherosclerotic lesions. Thus,
pre-existing cellular and humoral immunity to microbial
HSP60 may encompass pathogenetically relevant crossreactivity with HSP60 expressed on the surface of arterial
EC and leads to the initial inflammatory stage of atherosclerosis [10]. In addition, bona fide autoimmunity
triggered by biochemically altered autologous HSP60
may also occur [26]. Based on this concept, the “immunological hypothesis” of atherogenesis was formulated
[21]. Accordingly, the latter results strongly support
Virchow’s theory of atherosclerosis as a primarily inflammation-induced disease.
We recently had the unique opportunity to investigate
paraffin sections of early atherosclerotic lesions from
specimens secured by von Rokitansky in the middle of the
19th century, which were stored in the archives of the
Institute for Pathological Anatomy, Vienna. We used these
historical materials to confirm the presence and role of an
early inflammatory process in atherogenesis, and also
present methodological data on the successful use of
various modern immunohistochemical techniques on such
historical resources.
Materials and methods
Twelve atherosclerotic arteries, originally collected between 1827 and 1885, were selected from the Collection of
Pathological Specimens procured by Rokitansky and
thereafter stored at the Federal Museum of Pathological
Anatomy, Vienna, Austria (see Table 1). In the early 19th
Table 1 Origin of historical vessels from the Rokitansky collection
Museum archive number
Sex/age
MN1819a
MN3590
MN1943a
MN3854
MN2460a
MN1184a
MN1637a
MN1347a
MN1150
MN2918a
MN4207
MN2343a
Male
Male/51
Female/64
n.k.
n.k.
n.k.
Female/21
Male/47
n.k.
Male/43
n.k.
Male/49
n.k. not known
a
Specimens collected by Rokitansky
Year
1837
1878
1840
1881
1853
1828
1834
1831
<1827
1863
1885
1850
Tissue origin
A. brachialis
Aorta ascendens
A. lienalis
Aorta abdominalis
Aorta thoracis
A. lienalis
A. lienalis
A. lienalis
A. iliaca communis dexter
A. cerebri media sinister
A. cerebri media sinister
A. cerebri media sinister
98
century, specimens were stored in pure alcohol (“Weingeist”), and subsequently in “Kaiserling’s fluid” (750 ml
formaldehyde, 1,000 ml distilled water, 10 g potassium
nitrate, 30 g potassium acetate) [15]. In the beginning of the
20th century, Kaiserling’s fluid was replaced with lower
graded formaldehyde to preserve tissue to the present [3].
Historical human lymph node (1893) and human skin
(1849) proved to be unusable as positive control tissue as
either black deposits (lymph node) or section detachment
from the slides made successful processing impossible.
Formaldehyde (4% formaldehyde in phosphate buffered
saline, PBS, pH 7.2)-fixed recent samples of human skin,
tonsil, lymphnode, and arterial fragments were, therefore,
used for control purposes. For the final analysis, 3 of 12
vessels were excluded because of signs of autolytic
degeneration.
Historical and contemporary tissues were dehydrated
and paraffin embedded. Sections 5-μm thick were cut and
placed onto silane-coated slides. Sections were rehydrated
and the crucial step of antigen retrieval was performed
either with proteinase XXIV (P8038, Sigma, 1 mg/ml in
PBS, 5-min incubation) or microwave irradiation (750 W,
Table 2 Antibodies, optimized conditions, and results from recent control tissues
Code
Dilutiona Antigen retrieval
A 0452
1:50
Polyclonal Neo
Markers
CD 68 (macrophages) KP-1
Dako
RB-360-A
1:10
M0814
1:50
CD 68 (macrophages) PG-M1
Dako
M0876
1:50
CD 68 (macrophages) Kp-1
Neo
Markers
Dako
MS-397-S
1:30
M0851
1:50
A 0082
1:200
Antibody against
Clone
Source
CD 3 (T cells)
Polyclonal Dako
CD 3 (T cells)
Smooth Muscle Actin 1A4
von Willebrand Faktor Polyclonal Dako
S 100 Protein
4C4.9
HLA-DP, DQ, DR
CR3/43
(MHC II)
CD 25 (IL-2 rezeptoe) 4C9
Fibroblasts
TE-7
CD106 (VCAM)
1.4C3
CD 54 (ICAM)
54C04
CD 62-E (E-Selectin) G4
CD 62-P (P-Selectin) 1E3
Neomarkers MS-296-P
1:50
Dako
1:50
M 0775
Novo Cas- NCL-CD25- 1:50
tra
305
Cymbus
CBL 271
1:50
Neo
Markers
Neo
Markers
Novo Castra
Dako
MS-1101-S
1:5
MS-1094-S
1:10
NCL1:50
CD62E-382
M 7199
1:50
Collagen type I
Polyclonal Cedarlane
CL50111AP 1:60
Collagen type III
Polyclonal Cedarlane
CL50311AP 1:30
Collagen type IV
CIV 22
Dako
M 0785
1:25
Fibronectin
Polyclonal Dako
A 0245
1:100
Conjugate Positive control
tissue
Citrate
EnVision,
AP
Citrate
EnVision,
AP
Citrate
G. a-m.
AP
Citrate
G. a-m.
AP
Citrate
G. a-m.
AP
Citrate
EnVision,
AP
Citrate
EnVision,
AP
Proteinase XXIV or G. a-m.
EDTA
AP
Citrate
G. a-m.
AP
Citrate
EnVision,
AP
Proteinase XXIV or G. a-m.
EDTA
AP
EDTA
EnVision
AP
Citrate
EnVision
AP
EDTA
G. a-m.
AP
Proteinase XXIV
G. a-m.
AP
Citrate
G. a-rb.
488
Citrate
G. a-rb.
488
Citrate or n.p.
G. a-m.
AP
Citrate or n.p.
EnVision,
AP
– to +++: semiquantitative assessment of staining intensity
G. a-m. AP Goat anti mouse AP, n.p. no pretreatment, G. a-rb. 488 goat anti rabbit IgG, Alexa 488
Determined in pilot studies
a
Control
staining
Human tonsil
+++
Human tonsil
++
Human tonsil
+++
Human tonsil
+++
Human tonsil
++
Human tonsil
+++
Human tonsil
+++
Human tonsil
+++
Human tonsil
+++
Human tonsil
+++
Human breast
skin
Human tonsil
+++
Human tonsil
+++
Human tonsil
++
Human
lymphnode
Human breast
skin
Human breast
skin
Human kidney
+++
Human breast
skin
+++
+++
+++
+++
++
99
10 min) with citrate buffer (10 mM, pH 6.0) or EDTA
buffer (0.1 mM, pH 8.0). After a final washing step in PBS,
samples were treated with antibodies listed in Table 2. The
two-step indirect method yielded the best results of several
tested detection systems. Therefore, for immunohistochemistry staining, a goat anti-mouse immunoglobulin
(D0487, Dako, Glostrup, Denmark) or Envision polymer
(K4017, Dako), both conjugated with alkaline phosphatase, were used. Nuclei were stained with hematoxylin
(S3309, Dako).
Immunofluorescence detection was performed with an
Alexa488-labeled goat anti-rabbit immunoglobulin (A11008, Molecular Probes, Eugene, OR, USA). For nuclear
staining in immunofluorescence tests, diaminophenylindol
(D1306, Molecular Probes) was used. Controls were
performed with normal mouse serum (X0910, Dako),
Fig. 1 Immunohistochemical
staining on 5-μm thick paraffin
sections of arterial vessels collected in the 19th century by
Carl von Rokitansky. Positive
cells are visualized with an
alkaline phosphatase labelled
conjugate. * Arterial lumen:
a staining for CD3+ T-cells
(red); aorta ascendens, 51-yearold male died in 1878 (Protocol
Nr. MN3590 see Table 1).
Original magnification ×400;
b negative control—rabbit Ig
fraction; aorta ascendens,
51-year-old male died in 1878
(MN3590). Original magnification ×400; c Von Willebrand
factor (red); arterial intima, male
(age unknown) died in 1837
(MN1819). Original magnification ×200; d Negative control—
rabbit Ig fraction; arterial intima,
male (age unknown) died in
1837 (MN1819). Original magnification ×200; e Fibronectin
(red); arterial intima; male (age
unknown) died in 1837
(MN1819). Original magnification ×200; f Negative control—
normal rabbit serum; male
(age unknown) died in 1837
(MN1819). Original magnification × 200; g Collagen type
IV (red); arteria lienalis, 47year-old male died in 1831
(MN1347). Original magnification ×200; h Negative control—
mouse IgG1, arteria 47-year-old
male died in 1831 (MN1347)
normal rabbit serum (prepared in our laboratory) and rabbit
total immunoglobulin fraction (X0903, Dako) or appropriate mouse immunoglobulin isotype preparations, i.e.,
mouse IgG1 (X0931, Dako), mouse IgG2a (X0943, Dako)
and mouse IgG2b (X0944, Dako). Finally, slides were
mounted in Kayser’s glycerol gelatine (109242, Merck,
Darmstadt, Germany).
Microscopic analyses were carried out either with a
Nikon Eclipse E800 microscope or a Zeiss Axiovert 100M
confocal laser scanning microscope.
Results
Immunohistochemical and immunofluorescence analyses
using monoclonal and polyclonal antibodies recognizing
100
various cell surface markers, adhesion molecules, and
extracellular matrix (ECM) components were adapted and
successfully used for these studies. The T-cell surface
marker CD3, the endothelial marker von Willebrand factor
(vWF), and the epitopes of the extracellular matrix proteins
collagen type I, III, and IV and fibronectin were preserved
and, thus, could be visualized.
In two of nine vessels, CD3-positive cells could be
detected in atherosclerotic lesions (Fig. 1a). To exclude
unspecific staining, negative controls were also performed
showing no staining (Fig. 1b). None of the historical
vessels were positive for the macrophage surface marker
CD68, the smooth muscle cell antigen alpha-actin, the
dendritic cell marker S100, HLA-DR (MHC II), a fibroblast marker as well as the adhesion molecules CD62-P (Pselectin) or CD106 (vascular cell adhesion molecule-1–
VCAM-1) (see Table 3). In contrast, vWF was found in the
vasa vasorum of almost all vessels examined. However,
only a few specimens also showed a vWF positive
endothelium lining the main arterial lumen (Fig. 1c,d).
Fibronectin-expressing structures were present ubiquitously in almost all analyzed lesions, particularly in
adventitia and thickened areas of the intima (Fig. 1e,f). In
half of the arteries, collagen type IV-positive structures
were found in the vasa vasorum (Fig. 1g,h). A specimen of
the arteria lienalis additionally showed collagen type IVpositive filaments in intima and media. Collagen type I was
also detected in the adventitia and media of two vessels
(Fig. 2a). All atherosclerotic lesions examined showed
abundant collagen type III in the adventitia and media. In
contrast, collagen type III-positive fibres were rarely found
in the intima (Fig. 2b–d).
Discussion
Methodological aspects
Human pathological specimens have been collected and
stored in anatomical museums for centuries [15]. This large
pool of historical material could, in principle, serve as a
rich resource for various types of research. However, thus
far, only a limited number of methods were available for
the examination of such sensitive samples, and immunohistochemistry could not be successfully implemented
[15]. Our laboratory has extensive experience in this area
[19] and, in fact, introduced immunohistological techniques into paleontological research, thus creating a new
subdiscipline, paleoimmunology [20].
The arterial vessels examined derived from pathological
human samples collected by von Rokitansky and his
antecessor Wagner and successor Heschl between 1827
and 1885. Intriguingly, autopsy reports compiled approximately 170 years ago are often still available [15], a
remarkable sign of a well-organized storage system
throughout the centuries.
Implementing the application of modern immunological
techniques to atherosclerosis research has been a challenge,
as available supply of historical arteries was limited, and
some vessels had to be excluded due to autolytic degeneration. The historical control tissues, intended to be used
for establishing the assays, turned out to be unsuitable due
to black deposits of unknown origin. Additionally, some of
the chemicals used for tissue fixation and storage
conditions (exposure to light or air) might have altered
the specimens. The sensitivity of different epitopes to the
extraordinary extended fixation time (up to 17 decades) is
clearly distinguished by comparing the results found with
polyclonal and monoclonal antibodies, respectively. CD3,
vWF, collagen type I, collagen type III, fibronectin positive
Table 3 Expression pattern of different atherosclerosis markers on historical vessels from the 19th century Rokitansky collection
CD3 (T-cells)
CD68 (macrophage)
Clone KP1
CD68 (macrophage)
Clone PG-M1
SMA (alpha-Actin)
vWF
S 100-protein
HLA-DR (MHC II)
Fibroblasts
CD62-P (P-selectin)
CD106 (VCAM-1)
Collagen type I
Collagen type III
Collagen type IV
Fibronectin
MN 1819
MN 1150
MN 3590
MN 1943
MN 3854
G 2460
MN 1184
MN 1637
MN 1347
–
–
–
–
+
–
+
–
–
–
–
–
–
–
–
–
–
–
–
–
–
n.t.
–
–
n.t.
n.t.
n.t.
–
+++
–
–
–
–
–
+
+++
–
+++
–
+++
–
–
–
–
–
–
+++
–
+++
–
+
–
–
–
–
–
–
+++
–
+++
n.t.
++
n.t.
n.t.
n.t.
n.t.
n.t.
n.t.
n.t.
n.t.
n.t.
–
+++
–
–
–
–
–
–
+++
+++
+++
–
+++
–
–
–
–
n.t.
+
+++
–
++
n.t.
+++
–
–
–
–
n.t.
n.t.
n.t.
++
+++
n.t.
–
–
–
–
–
n.t.
n.t.
n.t.
+
+++
n.t.
+
–
–
–
–
n.t.
n.t.
n.t.
++
+++
– to +++: semiquantitative assessment of staining intensity
n.t. not tested
101
Fig. 2 Indirect immunofluorescence staining on 5-μm thick
paraffin sections of historical
human arterial vessels. *Arterial
lumen: a Collagen type I (FITC,
green) and nuclear staining
(DAPI, blue); adventitia of an
arteria iliaca (MN1150). Original magnification ×400; b Collagen type III (FITC, green);
arteria brachialis, male (age unknown) died in 1837 (MN1819).
Original magnification ×200; c
Transmission image from negative control for d; arteria brachialis, male (age unknown)
died in 1837 (MN1819). Original magnification ×200; d Negative control—rabbit Ig fraction;
arteria brachialis, male (age unknown) died in 1837 (MN1819).
Original magnification ×200
fibers and cells were all detected with polyclonal
antibodies, whereas, monoclonal antibodies generally
failed to detect these structures, with the exception of
collagen type IV, in historical tissue. In view of the unique
nonfibrillar procollagen-like structure of collagen type IV,
which includes several non-helical domains that are
susceptible to conventional proteases, the latter observation
was rather surprising.
It is, thus, probable that epitopes in tissue specimens,
especially after extended fixation time and harsh storage
conditions, deteriorate or are masked by chemical
processes such as methylen bridges formed between
reactive sites of tissue proteins. Thus, destruction or
masking of the epitope recognized by a monoclonal
antibody results in the lack of any signal. On the other
hand, the use of a polyclonal antibody greatly enhances the
possibility that at least one of the several epitopes these
immunoglobulins bind to has resisted deterioration.
Accordingly, the antigen retrieval procedure, has turned
out to be a crucial step. Finally, the optimal technique for
visualization of histological structures in historical tissue
samples, i.e., immunohistochemistry or immunofluorescence, must be determined for each antigen–antibody
system in pilot studies.
General aspects
It has long been known that both humoral and cellular
immune reactions take place in atherosclerotic lesions, but
it has not been clear whether these processes were primary
or secondary in nature. Already in the middle of the 19th
century, Carl von Rokitansky and Rudolf Virchow pointed
out an association of histologic signs of inflammation with
atherosclerotic lesions. Both described cellular inflammatory changes in atherosclerotic vessel walls, but while von
Rokitansky considered these changes secondary in nature,
Virchow supported their primary role in atherogenesis.
Atherosclerosis is a multifactorial disease, and advanced
lesions are by definition complex encompassing various
inflammatory and non-inflammatory hallmarks [28]. In
principle, one can distinguish three major types of this
disease, viz.
(a) Those that develop exclusively on a genetic basis and
for which valuable murine knockout models have been
developed [29], e.g., familial hypercholesterolemia
(FH) due to a heterozygous or homozygous deficiency
of the LDL receptor on one end of the spectrum.
(b) Mainly immunologically caused variants, i.e., allotransplant-associated atherosclerosis, are positioned at
the other end of the spectrum.
(c) Between these two extremes, the large majority of
atherosclerotic patients can be placed who develop
what we call poor man’s atherosclerosis, i.e., the form
that can afflict nearly everybody and that represents
killer number one in the developed world. Advanced
lesions of the latter type are characterized by the
presence of macrophages, smooth muscle cells, T cells,
dendritic cells, mast cells, fibroblasts, and abundant
102
collagenous and non-collagenous extracellular matrix
proteins. Endothelial cells in the atherosclerotic region
differ in their expression of adhesion molecules from
other, non-affected vascular territories [7]. Macrophages and smooth muscle cells possess non-saturable
scavenger receptors and are transformed into foamcells
that have the tendency to disintegrate and release their
lipid-rich content into the extracellular space where
cholesterol crystals may even be formed. Lymphocytes
are preferentially of the Th1 phenotype and include
both, effector and regulatory cells, as reflected by their
respective cytokine profiles [28]. B cells are scarce,
both in early as well as in late lesions [4, 7], but
antibodies against plaque antigens, inhibitory Fcreceptors, and cytokines are deemed to play a major
role in the advanced atherogenic process [30].
It now seems clear that inflammatory immunologic
processes on an appropriate genetic background are
operative in incipient atherogenesis, but the very first
pathogenetic event remains to be elucidated. In our
“autoimmune concept of atherogenesis” [21], we postulate
that classical atherosclerosis risk factors, such as hypertension, smoking, diabetes, the life-long infectious load or
an altered lipid metabolism first act as stressors of
endothelial and intimal cells leading to the expression of
HSP60 that then triggers adaptive and innate immune
processes that start the disease. With respect to the activity
of oxidized LDL as an endothelial stressor, it is of interest
that lipid-lowering statins have anti-inflammatory [31] and
immunosuppressive [32] effects. The initiating role of
adaptive immune mechanisms is, e.g., supported by the fact
that atherosclerosis can be induced in normocholesterolemic rabbits by immunization with recombinant HSP60
[24] and that the transfer of the disease in mice is possible
by both anti-HSP60 antibodies [33] or T cells [34].
Alternatively, components of the innate immune system
operative via the activation of Toll-like receptors (TLRs)
[35] or in the form of natural killer T (NKT) [36] cells may
initiate the disease.
Immunohistological studies in rabbits and humans have
identified activated T cells as the first intima-infiltrating
elements, clearly preceding macrophages and smooth
muscle cells [25]. The present results further support our
previous findings that CD3+ T cells are present in the
earliest phases of atherosclerosis and, thus, seem to agree
with Virchow’s view that inflammation plays a primary
role in initiating the atherogenic process rather than
Rokitansky’s concept assigning them a secondary role only.
This investigation of historical biological sources and
the application of modern technologies thereto has opened
new perspectives for the use of this valuable material for all
types of pathohistological studies.
Acknowledgements We want to thank Ruth Pfeilschifter-Resch
and the medical technologists at the Department of Clinical
Pathology in Vienna for preparing paraffin sections as well as
Ilona Lengenfelder for assistance in the preparation of figures and Dr.
Beatrix Patzak for consulting in sample selection. The Clinical
Department of Plastic and Reconstructive Surgery, University
Hospital Innsbruck, Austria and Dr. Felix Offner from the
Department of Pathology, County Hospital Feldkirch, kindly
provided recent control tissue. The authors want to acknowledge
the Austrian Science Fund (FWF-project no.14741 to G.W.) and the
European Union (Molecular basis of vascular events leading to
thrombotic stroke, MOLSTROKE; LSHM-CT-2004-005206) for
supporting the project.
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