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(Circulation. 1995;92:2876-2885.)
© 1995 American Heart Association, Inc.
Articles |
From the Second Department of Surgery (C.H., M.I., A.Z., E.W.) and Department of Urology (A.S., C.R., G.K., G.S.), University of Vienna, Austria.
Correspondence to Dr Christoph Holzinger, Allgemeines Krankenhaus Wien, Abteilung Herz- und Thoraxchirurgie, Währinger Gürtel 18-20, A-1090 Vienna, Austria.
| Abstract |
|---|
|
|
|---|
Methods and Results Leukocytes were quantified as reactive cells
per square millimeter in perivascular, interstitial, and
parenchymal tissue sections. Freshly isolated heart-tissue T cells
and peripheral-blood T cells from the same patients
were analyzed by triple staining and flow cytometry to identify
T-cell subpopulations as well as their states of differentiation
(expression of CD45RA and Leu-8 versus CD45RO) and activation (IL-2R,
IL-7R, very late antigen1, HLA-DR). All types of infiltrating cells
(T cells, B cells, macrophages, granulocytes) are increased in
hearts with IDC compared with normal hearts, but only CD8+
T cells and macrophages are increased relative to the other
leukocyte subpopulations.
CD45RO+/CD45RA-/Leu-8-
cells constitute the majority of heart-tissue T cells in both
normal hearts and hearts with IDC. Strikingly, hearts with IDC are
infiltrated by eightfold greater numbers of perivascularly located
IL-2R+ (26% of all T cells) and
CD45RO+activated memory T cells; moreover, in
contrast to normal heart,
40% of both CD4+ and
CD8+ heart-tissue T cells express activation
markers.
Conclusions Both normal hearts and hearts with IDC are populated by leukocytes. The quantitative increase in IDC, associated with a dramatically altered activation status of heart-tissue T cells, suggests a direct role of infiltrating leukocytes in the pathogenesis of IDC.
Key Words: cardiomyopathy lymphocytes genes cells
| Introduction |
|---|
|
|
|---|
Despite fundamental disagreement on many points, all of the cited reports share the observation that IDC is associated with an immunological process. Although this evidence essentially rests on analysis of peripheral-blood cells and largely unspecific investigations of the local infiltrate, the present study for the first time elaborates on the distribution and composition of cardiac tissue leukocytes, with the emphasis on T cells and their state of activation.
To determine which, if any, leukocyte subpopulations might have a
pathogenic role, we evaluated the differential distribution of
CD3+ T cells, their CD4+ and
CD8+
subsets, CD20+ granulocytes, CD67+ B cells,
and
CD14+ macrophages in normal hearts (n=4), hearts
with IDC (n=6), and hearts with ischemic congestive heart
disease (n=3) by use of a number of recently developed monoclonal
antibodies. In the next step, we addressed the state of differentiation
and activation of tissue T cells derived from normal hearts compared
with tissue T cells from hearts with IDC and the corresponding
peripheral-blood T cells from the same patients. It was
therefore necessary to develop a preparation technique for heart
tissuederived lymphocytes that allowed triple staining and
analysis by flow cytometry. Subsequent screening for
differential CD45RO (memory T cells), CD45RA (naive T cells), and Leu-8
(lymph nodehoming receptor) expression enabled clear phenotypic
differentiation between myocardium-specific tissue T
cells and incidental contaminating or passenger
peripheral-blood T cells; and triple staining for IL-2R
-chain (CD25) and ß-chain, IL-7R, VLA-1, and HLA-DR was
performed to evaluate their state of activation. By comparison with the
phenotype of peripheral-blood T cells of the
same patients, we analyzed whether the infiltrates were due to
unspecific influx or specific migration and whether T-cell activation
was local or systemic. Last, we addressed the question of whether
infiltration is acute or chronic.
| Methods |
|---|
|
|
|---|
Clinical Features
Criteria for diagnosing IDC were duration
of symptomatology,
gross enlargement of the heart relative to sex and age, congestive
heart failure (in the absence of significant coronary artery
sclerosis or valvular disease), and the microscopic findings of
myocyte hypertrophy, nuclear hypertrophy,
myocyte degeneration with myofibrillar attenuation, and
interstitial and perivascular fibroses. All patients were
in New York Heart Association functional class IV.
Echocardiography, chest radiograph, gated
radionuclide scan, and bilateral catheterization were
performed in all cases. All IDC patients had highly dilated ventricles,
as determined by left ventricular diastolic
dimension, a cardiothoracic ratio >0.55, a severely depressed ejection
fraction <15%, extremely poor contractility, and a
normal coronary angiogram. Myocarditis was excluded on the
basis of the Dallas criteria21 by histopathological
evaluation of the excised organs. The postmortem examinations showed
dilatation and enlargement of all chambers of the hearts. Valves and
coronary arteries were normal. The clinical history of disease
had lasted at least 8 months (mean, 13.4±7 months), and all patients
had been on the transplantation list for at least 4 months. All
patients were followed up on an outpatient basis but were advised to
stay in the hospital for cardiac recompensation. All patients showed a
gradual deterioration of myocardial function without signs of acute
myocardial degradation. All patients met the World Health Organization
criteria22 for the classification of IDC. Exclusion
criteria at histopathological examination were hypertension,
irradiation, pregnancy, endocrine disease, familial
cardiomyopathy, exposure to toxic agents, and
history of alcohol abuse or valvular disease. Patients with
ischemic congestive heart disease had survived at least one
myocardial infarction, had a severely reduced left
ventricular function, intractability of heart
ischemia due to surgical revascularization,
or percutaneous transluminal coronary
angioplasty as determined by coronary angiography. Control
tissues from clinically normal hearts were harvested from multiorgan
donors whose hearts were refused because of incipient coronary
plaque formation of the left descending coronary artery (n=2),
a bicuspid aortic valve (n=1), or logistic problems in the
transplantation hospital (n=1).
Tissue Preparation
Heart specimens were frozen rapidly in
isopentane in a liquid
nitrogen bath and stored in liquid nitrogen. Frozen sections were cut
into serial sections 4 µm thick, air-dried for 1 hour, and fixed
in acetone for 10 minutes at 4°C.
Immunoperoxidase Technique
Controls for each staining
procedure included irrelevant
immunoglobulins of the appropriate isotype and/or second-step
reagent controls without first antibody. Acetone-fixed cryocut
sections were preincubated in 10% normal goat serum (Gibco BRL Life
Technologies) for 30 minutes at room temperature, then incubated in the
first antibody (Table 1
; References 23 through 33)
either overnight at 4°C or 1 hour at room temperature, followed by a
PBS wash and incubation with biotinylated rabbit anti-mouse Ig
(Dakopatts A/S) and Strept-ABComplex/HRP (Dakopatts) developed in
3,3-diaminobenzidine. Counterstaining was performed with hematoxylin,
then sections were coverslipped in Aquamount (BDH Ltd).
|
Quantitative Analysis of Tissue T-Cell
Subpopulations
For enumeration of the various cell subpopulations,
immunolabeled serial sections were analyzed with a rectangular
ocular grid with a x40 oil objective. Approximately 100 fields within
heart specimens were randomly chosen, and the density of positive cells
was determined and expressed as the number of cells (±SD) per square
millimeter of cryocut sections 4 µm thick. Microscopic tissue
sections were reviewed with the sole purpose of characterizing the
inflammatory cell infiltrates as follows: (1) inflammatory cell type:
characterized and categorized by expression of various membrane
antigens recognized by monoclonal antibodies and (2) location:
perivascular (<30 µm from the endothelium),
parenchymal (defined as the presence of inflammatory cells between
myofibers without intervening fibrosis), and interstitial
(regions of coarse, patchy, confluent areas of fibrosis in areas of
fine interstitial fibrosis). Counted cells were quantified
as cells per square millimeter. Intravascular cells were also
analyzed, but because of the mechanical processes involved in
the various washing steps, their presence was eventually purely random
and the data inconclusive. Therefore, they were not included in the
study.
Cell Preparation and Enrichment of Heart-Tissue T
Cells
IDC tissue samples (50 to 100 g) were kept and minced into 1- to
2-mm3 fragments in PBS/heparin (Novo Nordisk A/S) and EDTA
(Sigma Chemical Co) to prevent blood coagulation. Small pieces were
then extensively washed by repeated intensive vortex treatment and
short sedimentation in PBS/heparin until tissue fragments became white
(light pink). Enzymatic dissociation and cell separation as previously
described34 were modified to create optimum conditions for
tissue T-cell enrichment. Heart fragments were incubated with a
solution of 200 U/mL type I collagenase (Sigma) and 100
µg/mL of DNAse type I (Sigma) in RPMI 1640 medium plus 10% FCS
(complete medium). Digestion overnight revealed a
homogeneous cell suspension without undigested larger
particles, suggesting that all heart-tissue T cells were
present in the suspension. Further enrichment, basically designed
to remove major and minor debris, was performed by filtration through
nylon wool and a 20% Percoll (Pharmacia) gradient
centrifugation. Cells from the pellet were then
adjusted to 1x107/mL and layered on top of a
discontinuous Percoll gradient containing 20%, 15%, 7.5%, and 5%
Percoll solutions, 2 mL for each fraction, in a 15-mL tube and
centrifuged for 20 minutes at 500g. Each Percoll
fraction was harvested separately, and high-density fractions
generated by 15% and 20% Percoll interfaces plus the pellet were
subjected to osmotic shock treatment in 10 mL distilled water at 4°C
for 2 minutes, followed by washing in complete medium. Killed cells
were removed by density centrifugation over a
Lymphoprep gradient (Nyco Med AS) at 37°C. Interphase cells were
collected and are referred to as enriched tissue T cells.
Three-Color Staining of Tissue and Peripheral-Blood
T Cells
Lymphoprep gradientderived cells were washed twice in
PBS
containing 0.2% NaN3 and 1% FCS. Aliquots of cell
suspension (100 µL) containing 1x105 cells were
incubated for 30 minutes at 4°C with FITC-, phycoerythrin-, and
peridinin-chlorophyll proteinconjugated monoclonal antibodies as well
as appropriate nonsense controls in all possible combinations. Two- and
three-color analyses were performed by flow cytometry
(FACScan, Becton, Dickinson & Co). Cellular debris and nuclei were
excluded by threshold and the conservative use of forward and side
scatters without altering the view of enriched lymphocytes and their
heterogeneity. Discontinuous Percoll gradient and
osmotic shock treatment were performed because
fluorescence-activated cell sorter (FACS)
analysis of the nonenriched population revealed intense
autofluorescence due to muscle and stromal cells. By using
multicolor systems (LYSIS Software, Becton Dickinson), we
were able to confirm that further enrichment had no effect on the
composition or the antigen expression pattern of heart-tissue T
cells. Osmotic shock treatment was used to kill nonlymphoid cells,
which, because of their inflexible cytoskeleton compared with T cells,
are unable to expand their surface effectively enough to survive.
Statistical Analysis
All values are expressed as
mean±SD. Significance of
differences was calculated with Student's t test. A value
of P<.05 was considered significant.
| Results |
|---|
|
|
|---|
Establishment of an Enrichment Method for Heart-Tissue T
Cells
Although immunohistochemical staining demonstrates whether a
certain antigen is expressed on the membrane of individual cells, it is
unsuited to analyze the percentage distribution of various cell
subpopulations, the presence of additional activation markers on the
same cell, or the intensity of antigen expression. Since the definition
of various cell subsets, and particularly any conclusion about their
state of activation, strictly depends on demonstrating more than one
antigen on the same cell, we established methods of cell separation
enabling analysis of cardiac T cells by three-color flow
cytometry.34 35
Our enrichment technique for heart-tissue T cells was described in detail in the "Methods" section. The crucial point was to consistently produce fragments small enough to allow extensive washing until the red color was gone. To validate our technique regarding peripheral-blood contamination, freshly prepared tissue and peripheral-blood T cells of the same patients were tested for Leu-8, CD45RA (characteristic of peripheral-blood T cells), and CD45RO (present predominantly on tissue T cells) expression.
One representative
triple-staining experiment is
depicted in Fig 1
, and Table 2
summarizes
the results for all 5 IDC patients. Fig 1
shows the histograms
of
CD45RO, CD45RA, and Leu-8 expression by CD4+ and
CD8+ tissue T cells and peripheral-blood T
cells in IDC. The purity of tissue T-cell suspension is best documented
by comparing the number of CD45RA+ and Leu-8+
cells of both origins. Whereas heart-tissue T cells
predominantly belong to the group of
Leu-8-/CD45RA-/CD45R0+
memory T-cell subset, most peripheral-blood T
lymphocytes express Leu-8 and CD45RA, and only a minor percentage
CD45RO. This phenotypic difference clearly demonstrates that
contamination of the tissue T-cell suspensions by
peripheral-blood T cells amounted to <10%.
|
|
Expression of Markers for Naive (CD45RA) Versus Memory (CD45RO) T
Cells and for Lymph NodeHoming Receptor (Leu-8) by
Peripheral-Blood and Heart-Tissue T Cells of IDC
Patients
The above experiments (Fig 1
, Table
2
) were also performed to
obtain data on the type of tissue T cells infiltrating IDC tissue. It
emerged that they belong predominantly to the group of primed T cells
known to constitute the "normal" tissue T-cell type in
nonlymphatic organs, which are also present in normal heart tissue,
albeit in significantly lower numbers. The demonstration of
significantly more CD4+/CD45RO+
helper/memory than
CD8+/CD45RO+ cytotoxic/suppressor memory
T cells among tissue T cells, which is in contrast to the ratio seen in
the peripheral-blood T-cell population of the same
patients, suggests a selection of newly incoming peripheral
T cells (Table 2
). The overlapping percentages of
CD45RA+/CD4+ (27.5%) and
CD45RO+/CD4+ (96%) indicate that
a population of about 20% of CD4+ tissue T cells coexpress
both differentiation antigens and, thus, reflect a transient state of
primary T-cell activation.
Whereas CD45RO expression was demonstrated
on almost all
CD4+ (96±7.2%) and most CD8+
(76.1±6.1%) tissue T cells in IDC, indicating that they bear
predominantly the memory T-cell phenotype, it was observed on
only a few CD4+ (27.8±2.2%) and CD8+
(33.2±4.7%) peripheral-blood T cells in IDC.
Conversely, peripheral-blood T cells were found to
include more CD4+/CD45RA+
(71±5.3%) and CD8+/CD45RA+
(94.5±6.7%) cells, compared with 27.5±4.3% and 28.2±4.7%
tissue T cells, respectively (Table 2
). Leu-8 (Mel-14)
expression
composed 11.1±3.7% of CD4+ tissue T cells,
12.4±1.2% of
CD8+ tissue T cells, 93.3±7.8% of CD4+
peripheral-blood T cells, and 78.6±6.6% of
CD8+ peripheral-blood T cells (Table 2
).
The relatively high percentages of Leu-8+ (mean, 23.5%)
and CD45RA+ (mean, 55.7%) T cells in tissue T-cell
suspensions, which were also confirmed in IDC tissue, suggest a
frequent ongoing process of lymphocytic infiltration in IDC tissue.
Expression of T-Cell Activation Markers by CD4+
T-Helper Cells and CD8+ Suppressor/Cytotoxic Heart-Tissue T
Cells and Peripheral-Blood T Cells in IDC
T cells can support or
suppress B cells and other lymphocytes, as
well as destroy target cells. To perform any of these functions, they
have to be activated. Therefore, we investigated the expression
of the T-cell activation markers IL-2R
-chain and
ß-chain, IL-7R, HLA-DR, and VLA-1. Intracellular signal
transduction induced by IL-2 is transmitted via high- or
intermediate-affinity receptors, indicating that the IL-2R
ß-chain has a role in driving the intracellular signal transduction
pathway. VLA-1 is the latest-emerging activation antigen currently
known30 36 and binds to connective tissue
compartments.
IL-7 is a potent stimulator of T-cell proliferation and, in contrast to
IL-2, is independent of any additional stimuli such as antigen
presentation.37 38 Reexpression of functional
IL-7R on tissue T cells was found to be associated with intense T-cell
infiltration.39 HLA-DR is an established T-cell activation
marker expressed by all activated T cells.32
Our
experiments revealed that all
immunohistologically analyzed IDC specimens
distinctly differ from normal heart in both the intensity and
distribution of activation marker expression. Fig 2
shows a representative experiment obtained with tissue
and peripheral-blood T cells from IDC patients.
Activation marker expression was found to be significantly increased on
both CD4+ and CD8+ tissue T cells (Fig
2
)
compared with peripheral-blood T cells. HLA-DR
expression was increased 4 times, VLA-1 >10 times, and IL-2R
ß-chain
3 times (Table 3
). Low-affinity
IL-2R
-chain was expressed primarily by the
CD4+ subset, without a significant difference between
tissue and peripheral-blood T cells (data not shown),
suggesting partial high-affinity IL-2R (heterodimer)
expression by tissue T cells. Similar results were obtained with an
anti-CD29 antibody recognizing all members of the integrin-ß1 family
(CD29high expression by CD4+ tissue T cells,
82.3%; by CD8+ tissue T cells, 87.3%; by CD4+
peripheral-blood T cells, 50.2%; and by
CD8+ peripheral-blood T cells, 45.7%).
Interestingly, IL-7R, usually a marker for
peripheral-blood T cells and found on tissue T cells
only in cases of extreme infiltration, as in transplant rejection, was
expressed by both CD4+ (17.3±3.2%) and
CD8+
(9.1±2.7%) tissue T cells.
|
|
Patterns of Leukocyte Subpopulations in Normal Heart Versus
IDC Tissue
The normal specimens exhibited small numbers of cells
(17±2.7/mm2) bearing the CD45 antigen, a panhematopoietic
cell surface marker, that were evenly distributed throughout the organ
(Table 4
).31 By comparison, all
CD45+ leukocyte subsets in all IDC tissues analyzed
(57.7±6.1 cells/mm2) were massively increased, accompanied
by an altered percentage distribution: Whereas the increase of
CD4+ T cells and CD20+ B cells was in
accordance with the overall increase in CD45+ cells, the
CD14 (monocytes/macrophages) and CD8 frequencies showed
additional fivefold and fourfold increases, respectively. This is
reflected in a shift of the T-cell ratio of CD4+ to
CD8+ (T-helper cells to cytotoxic/suppressor T
cells) from 4:1 in normal heart to 2.5:1 in IDC tissue.
CD67+ granulocytes were increased in absolute numbers but
decreased relative to the other leukocyte subpopulations (Table
4
).
|
| Demonstration of Significantly Increased CD45RO+ Memory T Cells in Various Compartments of IDC Tissue |
|---|
|
|
|---|
The number of cells expressing the above-mentioned markers
increased from the perivascular through the parenchymal to the
interstitial tissue areas (definitions in
"Methods" under "Quantitative Analysis of Tissue
T-Cell Subpopulations"). In normal hearts, CD45RA (naive T
cells) was expressed by 0.18±0.1 and Leu-8 by 0.35±0.2
cells/mm2 perivascularly, increasing through parenchymal to
interstitial (4.5±1.1 and 7.6±2.1 cells/mm2,
respectively). CD45RO (memory T cells) was expressed by 0.4±0.2
perivascular cells/mm2 and mostly by
interstitial cells (21±5.3 cells/mm2). If we
follow the relative percentages of both populations through the various
tissue segments, there was a shift away from
CD45RA+/Leu-8+ cells toward
CD45RO+ cells from perivascular (1:1) through
interstitial (3:1) to parenchymal (12:1) (Table 5
). By
comparison, IDC tissues overall showed
dramatically greater numbers of T cells, ranging from 3.1±0.9
perivascular to 134±27 interstitial CD45RO+
cells/mm2. Despite this sharp increase, the ratio of
CD45RO+ memory to CD45RA+ and
Leu-8+ naive tissue T cells in the three tissue
compartments was more consistent, the former outnumbering the
latter by about six times, suggesting a markedly intensified
lymphocytic influx compared with normal tissue (Table 5
).
Interestingly, the IDC samples showed only twice as many
CD45RO+ cells parenchymally as the normal hearts, whereas
the two other tissue areas exhibited a sixfold increase of memory T
cells compared with normal heart.
|
Expression of IL-2R and IL-7R in Various Compartments of Normal and
IDC Tissues
In normal tissues, expression of both IL-2R and IL-7R was
extremely low, constituting only 3.2% (0.47±0.12
cells/mm2) and 4.7% (0.66±0.13 cells/mm2),
respectively, of CD3+ T cells. In IDC tissues, IL-2R was
expressed by 18.4% and IL-7R by 8.6% of CD3+ cells. In
normal tissues, no IL-2R+ or IL-7R+ cells were
found perivascularly, and only small numbers in both the
interstitial and parenchymal compartments (0.3 and 0.17
cells/mm2, respectively) (Table 6
). By
comparison, the number of perivascular CD3+ cells in IDC
was four times higher, and 20% or 26% (0.2 or 0.3
cells/mm2) thereof reacted with antiIL-7R and
antiIL-2R, respectively, which confirms the finding that lymphocytic
traffic is intensified in IDC tissue (Table 6
).
Interstitial areas in IDC showed the most significant
increase in CD3+ cells, associated with strongly elevated
lymphokine-receptor expression (Table 6
). Predictably, the
percentage of IL-7R+ cells decreased from perivascular
(20%) through interstitial (16%) to parenchymal (5%),
indicating the pathway of lymphocytic traffic (Table 6
).
|
| Discussion |
|---|
|
|
|---|
The present study demonstrates that the overwhelming majority of
heart-tissue T cells are
CD45RO+/CD45RA-/Leu-8-
primed memory T cells. In the perivascular tissue compartment (ie,
where the phenotype of tissue T cells may be expected to come
closest to that of newly incoming T cells) of normal heart, only about
half of the CD3+ cells were CD45RO+ (Table
5
).
Compared with the phenotype of peripheral-blood
T cells of the same patients, the conclusion suggests itself that these
CD45RA- and/or Leu-8expressing perivascular cells are part of the
physiological lymphocyte turnover and not
immediately related to the localized tissue T cells. These patrolling
CD45RA+/Leu-8+ T cells advance as far as
the interstitium but are hardly demonstrable in the parenchyma. On the
other hand, the average influx of CD45RO+ cells into the
perivascular region is eight times as high in IDC (3.1
cells/mm2) as in normal heart (0.4 cells/mm2),
whereas the numbers of perivascular CD45RA+ and
Leu-8+ cells remain almost constant. Hence, this massively
stepped-up migration must be regarded as a selective process and,
in addition to the possibility of local T-cell proliferation, is one
possible explanation for the total T-cell increase in IDC tissue.
Together with the striking finding that a large number of perivascular
cells in IDC express the activation marker IL-2R, this means that
hearts with IDC, unlike normal hearts, are heavily infiltrated by
peripheral
CD45RO+/IL-2R+activated memory
T cells. Flow cytometry revealed that this cell pool includes both
CD4+/CD45RO+/IL-2R+
and
CD8+/CD45RO+/IL-2R+
cells in equal proportions.
The present study clearly demonstrates that the number of T cells is significantly increased in all tissue compartments of hearts with IDC and, even more importantly, that in IDC a great percentage of heart-tissue T cells express activation markers. The findings of increased numbers of CD45RO+, and especially of activated IL-2R+, tissue T cells in IDC compared with normal heart argue in favor of an ongoing immune response. A similar picture was obtained in a variety of human diseases, such as multiple sclerosis or rheumatoid arthritis,52 53 54 in which the majority of T cells are CD45RO+ and CD45RA-. Also, the interpretation that increased CD45RO expression in hearts with IDC reflects increased conversion from the naive to the memory state as a result of an ongoing immunologic response is fully consistent with patterns of tissue inflammation in leprosy55 : Whereas in tuberculoid leprosy there is an effective immune response associated with both accumulation of CD45RO+ memory T cells and strong antigen reactivity, lepromatous leprosy, which is associated with T-cell inflammation primarily of the CD45RA+ phenotype, is characterized by an inadequate immune response. A recent report showed that mild heart allograft rejection was associated with infiltration by predominantly CD45RA+ T cells, whereas in the event of moderate rejection, significantly more CD45RO+ infiltrating T cells were observed.56
The result that, with the exception of IL-2R
-chain on
CD4+ peripheral-blood T cells, all T-cell
activation markers used in the present study were (1) more
frequently observed on heart-tissue T cells than on
peripheral-blood T cells of the same patients and (2)
expressed in a similarly strong manner by both CD4+ and
CD8+ tissue T cells shows that the predominant
CD45RO+ memory T-cell phenotype demonstrated in
IDC tissue reflects ongoing activation rather than a resting
state after previous priming in situ. This conclusion is further
underlined by 43.55% of CD4+ and 15.1% of
CD8+ tissue T cells expressing IL-2R
-chain and
(50% of both tissue T-cell subsets expressing IL-2R ß-chain, in
the complete absence of any ß-chain expression by
peripheral-blood T cells. Similarly, both tissue T-cell
subsets reacted intensely with antiVLA-1 (46% and 41%,
respectively) in the absence of any significant amounts of
VLA-1+ peripheral-blood T cells. Since the
integrin-ß1 family members are receptors responsible for the
interaction with matrix compounds, it would appear likely that in IDC,
upregulated VLA-1 and CD2941 expression (data not shown)
reflects activation and increased attachment of tissue T cells to the
myocardium,36 41 although extracellular matrix
components have also been shown to modulate lymphocyte behavior.
To summarize, our findings (1) support the hypothesis that cell-mediated immunity has a role in myocyte degeneration, or destruction, in IDC and (2) argue in favor of an ongoing, chronic immune process in its pathogenesis. This is confirmed by the fact that the distribution of peripheral-blood T-cell subsets was not significantly different from that of tissue T cells, suggesting that tissue T cells from normal heart are identical to those involved in initiating and maintaining this process.
| Acknowledgments |
|---|
Received March 2, 1994; revision received May 17, 1995; accepted June 13, 1995.
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