Vol.2, No.1, 49-58 (2012) Open Journal of Immunology
http://dx.doi.org/10.4236/oji.2012.21006
Serological diagnostics of myocardium diseases
based on multivariate analysis of cardiotrophic
autoantibodies’ profiles
Olga Moiseeva1, Lubov Mitrofanova1, Elena Karelkina1, Dmitry Zverev1, Dmitry Lebedev1,
Serge Skurydin2*, Alexander Polet aev2,3
1V.A. Almazov’s Federal Center for Heart, Blood and Endocrinology, St. Petersburg, Russia
2Medical Research Center “Immunculus”, Moscow, Russia; *Corresponding Author:
3P.K. Anokhin Research Institute of Normal Physiology, RAMS, Moscow, Russia
Received 2 November 2011; revised 16 December 2011; accepted 30 January 2012
ABSTRACT
We analyzed profiles of Ig G a u toantibodies t o 1 6
cardiac specific proteins and their main immu-
nogenic region B-epitopes, in the groups of al-
ready verified cardiac pathology: acute and chro-
nic lymphocytic myocarditis, ST elevation myo-
cardial infarction, postinfarction remodeling of
myocardium, dilated cardiomyopathy and in heal-
thy controls along with patients, suffered from
gastritis (to evaluate immune response against
cross-reactive B-epitopes). AAB specific pat-
terns allowed us to distinguish cases among
themselves by means of multiparametrical ca-
nonical discriminant analysis in approximately
95% of cases. Positive predictive value in the
group of MYO reached 95%, in the STEMI—89%,
in the PIR—99%, in the DCM—99%, in the group
of gastritis—88%. Principal component analysis
of mentioned cardiac pathologies extended cur-
rent clinical knowledge of their immunopatho-
genesis. Obtained data markedly proved a us-
ability of serum AAB profiling for non invasive
screening, differential diagnostics and working
hypothesis composition.
Keywords: Autoantibody; Autoimmunity; B-Epitope;
Autoantibody Profiling; Cardiology; Principal
Component Analysis
1. INTRODUCTION
Alongside with direct damaging effect of infection or
toxic agent, autoimmune pathology seems to be impor-
tant in the pathogenesis of myocardium inflammatory
diseases. However our ideas about autoimmune patho-
genesis are based on indirect descriptive data only. Main
stream in current clinical research is to estimate onedi-
mensional laboratory marker or single clinical effect,
especially by means of multicenter clinical analysis, which
proves, for example, that the raised risk of dilated car-
diomyopathy development is accompanied with eleva-
tion of autoantibody (AAB) against beta1-adrenoreceptor
[1-3]. But there is always a problem: what is the cause-
effect relationship between the disease and accompany-
ing immunologic deviations, especially if to look at issue
from position of immunophysiology and natural immu-
noregulation [4].
Primary conception about the destructive role of any
autoimmune response was related to “horror autotoxicus”
concept by Paul Ehrlich [5]. However, the initial idea has
undergone an essential transformational challenge. The
basis of modern conception of immunity and autoimmu-
nity has been founded by Ilya Mechnikov, who argued
the immune system not to be designed for much defense
against microorganisms, but for dynamical maintenance
of “harmonious state” in the conditions of constant pres-
sure of Environment [6]. Development of Mechnikov’s
concept has led to understanding that one of basic func-
tion of immune system is rather Self-identifycation, Self-
maintenance, and Selfreparation throughout a life [7,8],
without focusing on antiinfectious protection. Really nor-
mal microflora of mucous membranes is involved in
human homeostasis and plays important role in many
physiologic functions. For example, saprophytes in the
gut take part in food digestion and utilization as well as
in production of vitamins [9]. Thus, it is evident, that the
immune system ignores normal microflora, and the pri-
mary function of immune system is not so much de-
voted to eliminate foreign genetic material [10] but to
maintain of basic homeostasis of the body [11-14]. Thus
autoimmunity can be surveyed, on the one hand, as the
important instrument of cell death or harmful metabolic
yields elimination from an organism, and on another
hand—as the instrument of immunochemical regulation

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 4 9-58
50
of homeostasis, alongside with neurologic and endocri-
nologic regulation [12,14-16].
All lymphocytes, matured in thymic cortex and me-
dulla—have undergone positive and negative selection,
and thus have become moderately autoreactive [17,18],
and autoreactive potential of the lasts is controlled by
tolerogenic mechanisms of antibody idiotypic networks
and Tregs [19-21]. Kovalyov was the first who described
the concordance between the levels of serum AAB and
the levels of binding autoantigens [22-24]. Accordingly
to Kovalyov, the interconnected cellular and humoral
immunity provides permanent balanced production of
great amount of natural immunoglobulin-like molecules
(i.e. AABs, BCRs and TCRs) in every health person for
clearance purposes [11], supporting immunochemical
homeostasis of the whole body. Proceeding from Kova-
lyov rule [24], AAB repertoire can serve as a mirror of
individual enzyme and protein reactivity, imprinting or-
gan- and tissue-specific disorders (feed-back principle).
In case of a disease, primary pathological process in any
organ may occur. It causes excessive cell death, active-
tion of necroptosis in spite of normal physiological apop-
tosis [25], and leads to excessive autoantigen presenta-
tion. Induction of destructive process in any organ or
tissue causes excessive cell death and consequent active-
tion of B- and T-lymphocytes by apoptic debris and
abundant exosomes [26], subclinical autoimmunization
[27,28] and secondary (compensatory) elevation of ap-
propriate AABs [11,29]. Such secondary autoimmune
responses are forwarded to clearance and reparation pur-
poses in the affected organ, in opposite to primary or
pathogenic autoimmune reactions based on genetic de-
fects of immunoregulation, which cause multiple devia-
tions [8]. In both cases (sanogenic as well as pathogenic),
individual elevation of serum AAB level is predictive
marker of organ specific cell death, autoimmunization,
and beginning (or present now) pathologic process [30].
The aim of investigating AABs’ profiles was to de-
velop the specific combination of markers to predict the
cardiac pathology and to esteem their contribution in the
immunopathogenesis.
2. MATERIALS AND METHODS
2.1. Patients
17 patients with acute and chronic lymphocytic myo-
carditis (the group Myo) were examined. The diagnosis
of myocarditis had been verified on the basis of Dallas
Criteria and immunohistochemical analysis of an endo-
myocardial biopsy at patients with recently sustained
heart failure and life threatening ventricular arrhythmia.
Multiple biopsies (4 to 6) were taken from the right ven-
tricle according to standardized techniques to confirm
lymphocytic myocardial infiltration. The immunohisto-
chemical analysis of biopsy specimens was made by
monoclonal antibody staining against CD3, CD4, CD8,
CD20, CD68 and HLA-DR (DAKO, Denmark). The
second group with ST elevation myocardial infarction
(STEMI) included 9 patients, who were inspected on 2 - 4
days of the disease. The average serum level of troponin
I was elevated significantly (32.3 ng/ml) in the STEMI
group. The third group included 8 patients with postin-
farction remodeling of myocardium (PIR). The fourth
group of dilated cardiomyopathy (DCM) included 11
patients.
When using coronarography, we had excluded all the
cases with heart failure of an ischemic genesis and myo-
carditis (according to endomyocardial biopsy) from this
group. The additional clinical aspect of patients with a
cardiovascular pathology is presented in Table 1. The
Table 1. Clinical profile of groups with a cardiovascular pathology. Abbreviations: FC—functional class, LAD—left atrial diameter,
LVEDD—left ventricular end diastolic diameter, LVESD—left ventricular end systolic diameter, IVSd—LV interventricular septal
wall thickness in diastole, PWd—LV posterior wall thickness in diastole, EF—ejection fraction by Simpson’s rule.
Myocarditis M ± SD
n = 17
STEMI M ± SD
n = 9
PIR M ± SD
n = 8
DCM M ± SD
n = 11
Age, years 37.8 ± 14.1 62.7 ± 9.4 49.9 ± 6.8 41.5 ± 15.3
Gender (m:f) 11:6 6:3 8:0 6:5
Body mass index, kg/m2 26.5 ± 3.3 28.7 ± 5.7 29.4 ± 4.8 28.6 ± 6.9
CHF FC—NYHA
STEMI group FC—Killip 2.2 ±1.2 1.0 ± 0.5 2.8 ± 0.6 2.5 ± 0.7
Ultrasound of the heart and vessels
LAD, mm 42.3 ± 6.5 42.0 ± 6.5 44.3 ± 5.4 46.8 ± 4.1
LVEDD, mm 57.8 ± 7.5 50.2 ± 7.1 67.4 ± 4.9 66.6 ± 7.0
LVESD, mm 43.9 ± 11.2 37.7 ± 9.3 56.7 ± 6.6 57.7 ± 8.3
IVSd, mm 10.3 ± 1.4 11.5±2.7 10.1 ± 3.4 10.2 ± 1.7
PWd, mm 9.7 ± 1.8 11.2 ± 1.0 9.7 ± 0.6 9.5 ± 1.5
EF, % 43.8 ± 17.8 45.7 ± 14.8 27.1 ± 7.6 28.7 ± 6.6

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 49-58 51
first control group (control group I, Health) contained 18
practically healthy volunteers (m = 8, f = 10; age 24 - 55
years; average age 39 years). The second control group
(control group II, Gastritis) contained 8 patients without
the signs of a cardiac pathology, but with chronic gastri-
tis, endoscopy confirmed (m = 3, f = 5; age 38 - 67 years;
average age 43 years).
2.2. ELISA Methods
Using standartized ELISA test systems for semi-quanti-
tave serum AAB evaluation (ELI-TEST group by Medi-
cal Research Center “Immunculus”, Moscow, Russia),
we defined individual normalized levels of organ marker
carditropic AABs against: cardiomyocyte cytoskeleton
antigen CoM-02, cytoplasmic antigen of cardiomyocytes
CoS-05-40, platelet antigen TrM-0.3, ANCA antigen,
NO-synthetase (NOS), angiostatin, cardiomyosin L, col-
lagen II and β2-glycoprotein I. All antigens were purified
in the MRC “Immunculus”, as described [31]. Besides
natural antigens synthetic 20-25-mer peptide fragments
(epitopes) of adenine nucleotide translocator-1 and -2
(ANT-1 and ANT-2), PAPP-A, р53, cardiac-type myosin-
binding protein C (MyBPC3), and an extracellular loop
of β1-adrenoreceptor, had been used (all produced by
“Peptide 2.0 Inc.” (Chantilly, USA)).
Mean individual immunoreactivity and normalized
serum AAB level to each natural antigen or synthetic
fragment was estimated as described [32]. Besides, we
analyzed specific multivariate features of AABs’ profiles
(integral autoreactivity pattern), i.e. spikes of normalized
AAB level, starting from mean individual immunoreac-
tivity level that was performed for every antigen to pro-
duce the profile [4,33,34].
2.3. Statistics
The statistical analysis of the received data was made
with nonparametric statistical methods (Mann-Whitney
test, Chi-square test, Kruskal-Wallis test) and methods of
multivariate analysis (principal component analysis PCA,
canonical analysis CA, discriminant analysis DA) with
the usage of software package Statistica 8.0. Exploratory
application of PCA made it possible to transform sig-
nificant correlations of numerous AAB content to in-
duced variable format (principal components), that had
facilitated a visual estimation of these AAB deviations
and had allowed to discover linearly invisible factors,
that did not exist before [33,35,36]. Discrimination rules
had been produced to analyze specific features of AABs’
profile of each patient. Canonical functions along with
PCA had been used to estimate visually peculiarities of
each patient distribution in the uniform coordinate frame
in which different nosology forms revealed their simi-
larities and differences.
2.4. Ethics
The study was approved by the Local Ethic Commit-
tee of V.A. Almazov’s Federal Center for Heart, Blood
and Endocrinology.
3. RESULTS
3.1. Serum Levels of Troponin I and IgG
Autoantibodies to p53, to ANT-1, to
MyBPC, to Cardiomyosin L and to
CoS-05-40 at Patients within MYO,
STEMI, PIR and DCM Groups
Significantly Differ
Comparative clinico-immunological analysis of patients
with myocarditis showed the raised troponin I serum
level (the marker of myocardial leison) to be associated
with elevation of AAB to protein р53 (rs = 0.651; p <
0.01). Along with it, higher troponin I level, 12.3 ± 3.9
ng/ml against 2.0 ± 3.5 ng/ml (p < 0.05) in other cases,
was determined at patients with raised AAB to protein
MyBPC3. Elevated histological ratio of cardiomyocyte
necrosis in myocardium specimens had been closely re-
lated to elevated AAB to CoS-05-40 (χ2 = 6.52; р < 0.05)
and to cardiomyosin L (χ2 = 6.52; р < 0.05).
AAB profiling of patients within MYO, STEMI, PIR
and DCM groups had shown significant intergroup dif-
ferences of AAB distribution for cardiomyocyte cytos-
keleton CoM-02 antigen (χ2 = 21.7; p < 0.01) and car-
diomyosin L (χ2 = 21.3; p < 0.01). In details, for the
cases in STEMI and MYO groups the raised profile was
significantly higher, than in the control groups, whereas
for patients with PIR and DCM it essentially did not dif-
fer from normal controls. Direct correlation dependence
between the level of AAB towards main immunogenic
region of ANT-1 and ejection fraction (rs = 0.362; p <
0.05) was established. The similar was also found be-
tween the level of AAB to a fragment of an extracellular
loop of β1-adrenoreceptor and the left ventricle dimen-
sions: LV EDV (rs = 0.404; p = 0.02) and LV ESV (rs =
0.436; p = 0.013).
3.2. Linear Differences of AABs Are Not
Specific for Each of Described Cardiac
Pathology, while AAB Profiling Reveals
Intergroup Differences
Despite intergroup linear differences (Kruskal-Wallis
test, p < 0.05; Median test, p < 0.05) (Figure 1), our at-
tempts to discriminate blindly the inspected patients to
one of the 5 clinical groups (myocarditis, acute myocar-
dial infarction, postinfarction remodeling, chronic gastri-
tis and practically healthy cases), only on the basis of
absolute values of a single serum antibody, had appeared

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 4 9-58
52
ineffective. During the same time, analysis of the indi-
vidual profiles, previously described in [4,12], having
reflected relative interconnections in a content of nu-
merous AABs (the pattern of integral autoreactivity of a
patient), had allowed us to discover significant inter-
group differences for several antigens. The cases of each
group possessed their representative AABs’ profiles,
which had made it possible to significantly distinguish
patients among themselves by means of canonical dis-
criminant analysis (Figure 2).
Figure 1. AABs’ profiles, appropriate of different clinical groups (based on average values).
Figure 2. Map of projections of canonical factor loads of cardiac patients on the axes of canoni-
cal roots (based on AABs’ profiles and their correlations).

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 49-58 53
3.3. Multivariate Analysis of AABs’
Distribution (PCA, CA, DA) at Cardiac
Pathology Provides Additional
Information about Its
Immunopathogenesis
Using a multiparametrical factor analysis for study of
AABs’ profiles of patients in the STEMI group, some re-
gularity in profile distribution had been noted (Tables 2
and 3). In the structure of the first principal component at
patients with acute myocardial infarction the basic con-
tribution was made by variance in content of AAB to
ANT, MyBPC3, cardiomyosin. The factor loadings by
marker AABs to ANCA, β2-GP I and TrM-0.3 had ap-
peared to become essential less (being loaded of the 2nd
and 3rd factors). Marker AAB to β1-AR loaded the struc-
ture of the 5th principal component.
Marker AABs’ profile, specific for MYO, looked a little
Table 2. The contribution of some AABs in the structure of principal components of cardiac clinical groups. Digits designate se-
quence numbers of corresponding principal components. For simplification of interpreting only the first 7 principal components from
each group are chosen. The criterion of variable inclusion (i.e. marker AAB) in a certain principal component is considered, if factor
weight of a variable AAB for this component exceeded more than 0.7. Empty cells mean the presence of minor residual correlation or
insignificant factor weight of AAB outside of 7 principal components. For simplification of interpreting, each cell is coloured in a
spectrum range: from red (the less significant 7th component) to violet (the most significant 1st component).
AAB to AG
Health Myocarditis
Acute myocardial
infarction
Postinfarction
remodeling
Dilated
cardiomyopathy Gastritis
MyBPC3 2 1 3 6 3
ANT-1 (epitope VDP) 4 1 5 5 3
ANT-2 (epitope EGS) 7 1 3 3
β1-AR 1 3 5 4 5
p53 5 7 2 1
Cardiac myosin L 2 1 1
β2-GP 3 2 3 3
CoМ-02
1 1 2 1
СоS-05-40 3 2 4 1 4
Cardiac myosin 1 1 1 3
TrM-0.3 5 5 3 3 2 4
ANCA 2 4 2 4 5 5
NOS 6 6 5 2
Angiostatin
7 1 2
РРАР-А 3 4 2 7 1
Collagen II 1 2 6 2 2 2
Table 3. Cumulative eigenvalues for denoted principal components.
Principal
components Health Myocarditis
Acute
myocardial
infarction
Postinfarction
remodeling
Dilated
cardiomyopathy Gastritis
1 22.91% 37.21% 39.24% 40.48% 46.16% 49.01%
2 39.80% 58.62% 59.84% 65.42% 63.31% 65.64%
3 54.62% 69.29% 73.47% 85.52% 74.37% 77.96%
4 64.42% 76.34% 81.30% 95.05% 82.26% 86.58%
5 73.45% 82.32% 87.82% 98.18% 89.20% 93.22%
6 81.48% 87.54% 93.45% 100.00% 94.18% 96.95%
7 86.89% 91.10% 97.38% 96.66% 100.00%

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 4 9-58
54
differently. The structure of the first principal component
was loaded by AAB to cardiomyosin and to cardiomyo-
cyte cytoskeleton CoM-02 antigen. Marker AAB to β2-
GP I took a place in the second principal component, as
well as AAB to collagen and to CoS-05-40 antigen.
Marker AAB to β1-adrenoreceptor also enlarged their
weight, being risen from the 5th component in the
STEMI group to the 3rd component in MYO. At the
same time AAB to ANCA, whose modifications were
evidently expressed in STEMI, had achieved the low
factor importance at the myocarditis group, being taken a
place of the 4th principal component.
In the PIR group the first principal component was
presented by AAB to angiostatin and to CoS-05-40 anti-
gen. The structure of the second principal component
was loaded by AAB to collagen, PAPP-A, NOS and my-
osin light chain. The structure of the third principal
component of PIR was loaded by AAB to ANT, MyBPC3,
cardiomyosin and to platelet TrM-03 antigen.
In the DCM group the first principal component was
presented by AAB to a myosin light chain. The structure
of the second principal component was filled with AAB
to apoptosis regulator p53 and platelet TrM-0.3 antigen.
Factor weight of AAB to ANT, MyBPC3 and PAPP-A
had appeared to be the least in the DCM group.
At visual analysis of AABs’ profiles of various forms
of a cardiac pathology, some general lines in allocation
of AAB peaks had been noted. It is worth saying, that
factor similarity of group pairs: MYO and STEMI, PIR
and DCM, was discovered. Analysis of principal com-
ponent allocation had allowed us to differentiate the
specified clinical groups by means of several marker
AABs (Ta b l es 2 and 3). In other words, if not mention-
ing mutual immunopathogenic load of AABs, the basic
discriminants in the groups of MYO and STEMI had
appeared to be AABs towards MyBPC3, ANT, ANCA,
CoS-05-40 and to collagen II. The groups of PIR
and DCM were differentiated by AAB to plasminogen,
NOS, CoM-02, MyBPC3, ANT-2, β2-GP. The gastritis
group differed from the control group of healthy donors
by AAB to cardiomyosin L, cardiomyosin, plasminogen,
β1-AR, p53, CoM-02, ANT-2, ANCA.
At discriminant analysis of all groups simultaneously
among themselves there were discovered significant de-
viations in content of several AABs, whose had been
arranged in order of decreasing their significance level
p(DA, F = 5.79, p < 0.006): TrM-0.3, collagen, PAPP-A,
p53, angiostatin, β1-AR, CoS-05-40, β2-GP, CoM-02,
NOS, ANCA, cardiomyosin L.
3.4. AAB Profiling Provides Extra Help for
Making Diagnosis of Cardiac Pathology
By means of AABs’ profile usage it had become pos-
sible to confirm the diagnosis or to assign a patient to a
control health group in almost all of the cases. Usage of
discriminant analysis made it possible to diagnose cor-
rectly the nosology in 95% of cases, varying among the
groups from 88% (gastritis) to 100% (myocarditis) (Fig-
ures 2 and Table 4).
For application and choice of maximum value of each
of 6 discriminant functions (diagnosis establishment), we
had been discovered the conforming quotients and con-
stants. However for an approximate assessment of a state
of the patient (Figure 2) and making the provisional di-
agnosis, it was possible to take advantage of graphic
construction with the usage of canonical functions.
Let’s examine two specific cases.
Case Report 1. The patient M. of 32 years (f), was
hospitalised with complaints of sharp weakness and
tachycardia at the minimum physical effort and at rest.
Two months ago she had overcome a viral infection. At a
transthoracic echocardiography the dilatation of left
chambers of the heart with depression of global contrac-
tility and rising of pulmonary pressure was revealed. In
Table 4. Matrix of discriminant classifications for 6 clinical groups. In the first two columns there are primary clinical classifications,
in remaining columns—the predicted classifications via AAB profiles.
Percent
Correct
Health
(n = 20)
Myocarditis
(n = 18)
Acute myocardial
infarction (n = 8)
Postinfarction
remodeling
(n = 5)
Dilated cardiomyopathy
(n = 14)
Gastritis
(n = 8)
Health 95 18 1
Myocarditis 95 1 18
Acute myocardial
infarction 89 0 8 1
Postinfarction
remodeling 100 0 5
Dilated cardiomyopathy 100 0 13
Gastritis 88 1 7
Total 95 20 18 8 5 14 8

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 49-58 55
addition, arisen blood levels of troponin I (15.2 ng/ml),
C-reactive protein (6.13 mg/l) and NT-proBNP (1996
pg/ml) was found. Coronarography showed coronary ar-
teries to be without indications of any atherosclerotic
lesion, blood supply was not disturbed. Pressure in the
right ventricle was of 53/12 mm Hg. Necrosis of car-
diomyocytes with lymphocytic and macrophage infiltra-
tion was determined, along with perimuscular fibrosis,
HLA-DR-antigen expression on endothelium and on
infiltrating cells at endomyocardial biopsy. Magnetic
resonance tomography had confirmed diffuse myocardial
changes of an inflammatory character. There were no
PCR signs of gene elements of adenovirus, enterovirus,
HHV6, Parvovirus B19, CMV, EBV in situ in any biopsy
samples of the myocardium. During examination of
AABs’ profiles, some AAB rising towards peptide frag-
ments of Adenine nucleotide translocator 1 and 2, р53,
cytoplasmic antigen of cardiomyocytes CoS-05-40, car-
diomyosin L and platelet antigen TrM-0.3 was observed.
Case Report 2. Patient T., 27 years old (m), was hos-
pitalised for percutaneous CARTO-guided catheter ra-
diofrequency ablation of ventricular arrhythmia. Ven-
tricular disturbances came to be evident within last 2
years. Medicamental antiarrhytmic therapy was without
any essential effect. The patient noted transient loss of
consciousness. At Holter monitoring there were Regis-
tered right ventricular extrasystoles up to 16708 times
per day. The dimensions and contractility of the heart
were without essential deviations from norm at a trans-
thoracic echocardiography. Necrosis of cardiomyocytes,
perivascular fibrosis, edema of myocardial stroma, lym-
phocytic infiltration and expression of HLA-DR on cells
of an inflammatory infiltrate and an endothelium were
observed at an endomyocardial biopsy. There were no
any morphological signs of arrhythmogenic cardiomyo-
pathy and amyloidosis. It was found, that enterovirus
capsid protein VP1 had been expressed in myocardium
via immunohistochemistry. There were found arisen AAB
levels towards peptide fragments of Adenine nucleotide
translocator 1 and 2, р53, myosin binding protein MyBPC3,
cardiomyocyte cytoskeleton antigen CoM-02, platelet an-
tigen TrM-0.3, ANCA antigen, cardiomyosin L and β2-
glycoprotein I.
4. DISCUSSION
Nowadays diagnostics of inflammatory diseases of a
myocardium represents essential difficulties. In view of
lack of pathognomonic symptoms, and also low sensitiv-
ity and specificity of the majority of laboratory tests [37,
38] often there is a necessity of morphological refine-
ment of the disease diagnosis. At the same time, for myo-
carditis intravital diagnostics it is used from 4 to 6 biop-
tate specimens, and the standard pathoanatomical re-
search to make a diagnosis of myocarditis, assumed tak-
ing not less than 17 samples (which were histology- cally
proven in 80% of cases maximum) [39,40]. As a result,
the sensitivity of endomyocardial biopsy at use of one
biopsy specimen achieves 25%, and it is not elevated
above 50% at use of 4 - 5 biopsy specimens. The raised
level of troponin I as a marker of myocardial damage, is
determined at 35% of patients with myocarditis and is
characterised by high specificity (at about 89%), but low
sensitivity (about 34%) [41]. Therefore, normal level of
troponin or creatine kinase MB-fraction does not permit
to exclude the diagnosis of myocarditis [41]. Low speci-
ficity and rather high sensitivity characterise scintigraphy
of a myocardium by gallium-67 and by antimyosin AB,
labeled with indium-111 [42]. Last years the tomography
is suited for non-invasive diagnostics of inflammatory
diseases of a myocardium with gadolinium contrasting
amplification, which allows at usage, at least, of 2 to 3
criteria (LGE, EGE, T2-WI) to diagnose myocarditis
with sensitivity of 67% and specificity of 91% [43].
The present results show an effectiveness of immuno-
chemical methods of diagnostics in cardiology. Without
any detailed explanation of predicted role of AABs in
immunopathogenesis of one or another cardiac nosology,
whose load can be neutral, pathogenic or sanogenic, the
variation of serum content of principal AABs is often the
earliest and specific marker of any chronic pathologic
process [30].
Recently undertaken attempts to use the titers of one
or another “carditropic” AAB for diagnostics of myocar-
dities have appeared not quite successful. For instance, it
was shown, that raised AAB titers to sarcolemmal and
myofibrillar proteins of cardiomyocytes were found not
only in 12% - 75% of patients with myocarditis and di-
lated cardiomyopathy, but also in 4% - 34% of healthy
persons [44-46]. Moreover, inflammatory diseases of a
myocardium can reveal at normal level of some “car-
ditropic” AABs [47-50]. Therefore interpretation and
usage of AAB titers would become speculative and non
evident until it is based only on laboratory referential
“averaged norms”. The data sets with similar medians
but with different dispersions impose greatly on AABs’
profile formation, without any appeal to mean laboratory
norms. In this context we would like to cite Meroni
(2007): “…The initial paradigm ‘one autoantibody for
one disease’ does not appear to be useful any longer. An
autoantibody profile does seem to offer more diagnostic
and prognostic power than the determination of single
autoantibody specificity. The consequence is the use of
new assays to detect different autoantibodies” [34]. Backes
C. and other write: “Instead of allocating single antigens
to a specific group of diseases and even to a specific dis-
ease, it appears more appropriate to allocate seroreactiv-
ity patterns” [29]. This idea of autoantibody profiling is
shown by us and others [29,34].

O. Moiseeva et al. / Open Journal of Immunology 2 (2012) 4 9-58
56
As the data in our work conveys, only such multi-di-
mensional approach provides high specific and sensitive
discrimination of serums not only of cardiologic patients
from serums of healthy persons, but also to differentiate
various forms of a cardiac pathology and reveal molecu-
lar peculiarities of their immunopathogenesis.
It is worth saying, that the first and the second prince-
pal component of each nosology suit common knowl-
edge about its immunopathogenesis (Table 2). For ex-
ample, it is well known about the principal role of my-
osin lesion in MYO, ATP deficiency in STEMI, collagen
deposition in PIR, genetic defects of myosin in DCM.
Moreover, analysis of further components gives valuable
information about hidden processes of immunochemical
metabolism (even in norm), which needs further investi-
gation.
We suppose, that increasing the number of observa-
tions in a combination of usage of new perspective anti-
gens will allow us to improve diagnostic value of dis-
cussed approach and to transform it from being a con-
forming method to become a highly informative and
quite trivial differential-diagnostic laboratory procedure,
along with assistance for working hypothesis composi-
tion.
Our findings demonstrate that circulating AABs and
their profiles accompanying to several forms of cardiac
pathology can be considered as a marker of developing
disease, even not autoimmune by its origin, but with its
own component weight of autoimmunity.
5. ACKNOWLEDGEMENTS
Authors thank Kilikovsky V.V. (RSMU, Moscow) for fruitful com-
ments on usage of statistical methods and reviewing of manuscript.
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