Memory T cells are necessary for development of the immune response and represent one of the most numerous population of human T lymphocytes. On the contrary, suppressive regulatory T cells (Tregs) may terminate the immune response and help to maintain tolerance to self-antigens. These important groups of cells are consisting of different subpopulations and retaining throughout life. However, today there is yet no clear understanding of how the relations between these two groups of cells are formed. In this work we consider possible ways of development and maintenance of CD4+ T cell memory and role of Tregs in these processes. Mechanisms of a differentiation of memory T cells, Tregs and recently described memory Tregs are discussed. The functional and genetic characteristics of these cells are compared. Division of cells according to the functional profile allows drawing parallels between memory T cells and Tregs. These two groups are consisted of central circulating populations (Tc), effector which can migrate toward specific tissues (Te) and tissue-resident cells (Tr), which are staying in peripheral tissues. The similar structural organization of Tregs and memory T cells, existence of transitional forms of tissue-resident Treg subpopulations with properties of memory cells assumes existence of close interrelation between these groups of lymphocytes. The conversion of CD4+ memory T cells into FoxP3-expressing Tregs is one of possible mechanisms of communication between these two groups. The memory Treg-cells with T cell and memory Treg-cell properties can represent a transitional stage of differentiation. On the other side, Treg cells can differentiate independently of memory T cells and accumulate during life in the form of memory Treg cells. The supressor function of Tregs is also necessary as well as function of memory T cells to develop the immune response. It is possible, that a subset of Treg cells undergoes selection in thymus and constitutively express TCR-receptors having affinity with peripheral tissues. Further, these committed cells can be settled into tissues and become tissue-resident Treg cells which maintain regional T cell memory. Tregs can represent the “mirror image” of the structural organization of memory T cells, but with the return sign – the sign of suppression. The quantitative ratio of Tregs and memory T cells (CD4+CD45RO+CD25hiFoxP3+/CD4+CD45RO+CD25-FoxP3-), perhaps, is important criterion for functional assessment of immune system. The balance between these functionally opposite cell subsets has to provide stable functioning of immune system.

At the present time, a broad spectrum of CD8+ T lymphocyte subsets is revealed, including naïve cells, memory cells and regulatory subpopulations. Along with cells with high cytolytic activity, some subsets with marked regulatory activity were found there. Each subpopulation is characterized by a set of produced mediators, surface and intracellular markers allowing to suggest their differential in vivo functional activity. The present review article proposes a classification of CD8+ Т cells which takes into account their morphological and functional features. According to conventional view, the CD8+ Т lymphocytes is a cell population exhibiting high cytotoxic ability which is of critical significance in pregnancy, under the conditions of semi-allogenic fetal cell invasion into the endometrium. The fraction of CD8+ T cells is rather high in decidual structures. The review discusses the known mechanisms of differentiation regulation, selective migration and activity of CD8+ T cells in decidual membrane and placenta in the course of pregnancy. Perforine and granzyme are the main cytotoxicity factors of CD8+ Т cells. IL-2, IL-5, IL-13, IFNγ, IL-17, TGF-β and IL-10 cytokines are considered regulatory mediators of CD8+ cells. To induce the effector properties of CD8+ T cells, an antigenic stimulation is required, which is provided by interactions between the CD8+ Т cells and activated CD4+ Т cells or dendritic cells, cytokine effects. Specific differentiation of the CD8+ T cells is determined by differences in microenvironvent. In the course of pregnancy, accumulation of CD8+ Т cells is observed in decidual membrane, but their phenotype and functional properties differ from CD8+ Т cells in peripheral blood. At present time, the mechanisms of selective CD8+ T cell migration to decidual membrane are studied. These events are suggested to be mediated by means of CXCR3 and CCR5 chemokine receptors, IL-6 and IL-15 cytokines. The features of CD8+ Т cell activities, and production of some cytokines, e.g., CSF2, IFNγ, IL-1β, IL-2, IL-6, IL-8,IL-10, IL-12 and TNFα in decidual membrane and is of critical significance for effective invasion of trophoblast cells. In turn, the trophoblast and placental cells promote development of regulatory CD8+ Т lymphocytes in decidual membrane, being able to induce CD8+ T cell apoptosis in decidual membrane. Hence, interaction between the maternal CD8+ T cells and trophoblast in the area of uterine-placental contact is an important link during development of immunological tolerance in the maternal/fetal system.

The purpose of the present study was to specify a role of inflammatory mediators in pathogenesis of various types of anemia in pregnant obese women. We determined IL-1, IL-6, TNFα, C-reactive protein and hepcidin concentrations in blood serum of pregnant women with obesity depending on the type of anemic syndrome, either iron-deficiency anemia, or anemia of chronic diseases. We showed that the content of IL-6 in blood of the obese women exceeds the value of this index in healthy pregnant women (p < 0.05), and it does not depend on the presence and type of anemic syndrome. We found that the C-reactive protein concentration in pregnant women with obesity is higher than reference values (p < 0.05). Moreover, the contents of C-reactive protein in blood serum of pregnant women with anemia of chronic diseases is significantly higher (p < 0.05) than in women with iron deficiency anemia. Hepcidin concentration in blood of pregnant women with obesity and anemia of chronic disease was a specific feature: its content was significantly higher than in healthy pregnant women (p < 0.05), or in pregnant women with anemia-free obesity (p < 0.05). Hepcidin levels also exceeded 2-fold its contents in serum from pregnant women with obesity and iron deficiency anemia (p < 0.05). We have found that only pregnant women with obesity and anemia of chronic diseases have shown a positive correlation between the concentrations of C-reactive protein and blood levels of hepcidin (r = 0.733, p < 0.05), or IL-6 (r = 0.679, p < 0.05).
The discussion concerns potential mechanisms of evolving anemia of chronic disease combined with subclinical inflammation in pregnant women with metabolic disorders. We conclude that a combination of obesity with gestational diabetes is a risk factor of anemia of chronic diseases in pregnant women. Development of an algorithm for differential diagnosis of iron deficiency anemia and anemia of chronic diseases in this cohort of patients is advisable for future studies in the area.

Specific antibodies against environmental chemical gene toxicants and endogenous steroid hormones are shown to modulate concentrations of these compounds in blood serum and their biological effects in experimental models. However, probable hazards of such antibodies in human teratogenesis are still unknown. In particular, potential correlations between specific serum antibodies, sex hormone levels in pregnant women, and congenital malformations in newborns are not clear. The aim of this study was to identify possible associations between occurrence of antibodies to benzo[a]pyrene, estradiol and progesterone (Bp, Es and Pg, respectively), and congenital malformations, and effects of these antibodies upon Es and Pg concentrations in blood serum of pregnant women. We have included into the study 182 women with normal pregnancy and 101 females with congenital malformations of fetus. A non-competitive solid phase immunoassay was performed using Bp, Es and Pg conjugated to bovine serum albumin as antigens. Es and Pg serum concentrations were measured using immunoassay test-systems of “Immunotech” (Moscow). Results: strong positive correlations were revealed between the levels of studied antibodies in the both groups. High IgA-Bp/IgA-Es (> 3) and IgA-Bp/IgA-Pg ratios (> 3) were associated with congenital malformations (OR = 2.2, p = 0.013 and OR = 6.8, p < 0.0001). Positive correlations were revealed between Pg/Es and IgA-Bp/IgA-Es (rS = 0.62, p < 0.0001), and IgA-Bp/IgA-Pg ratios (rS = 0.77, p < 0.0001) in cases with inborn malformations. Similar correlations were found for the women who had normal pregnancy (rS = 0.4, p = 0.0001, and rS = 0.23, p = 0.026, respectively). The Pg/Es proportion correlated with IgG-Bp/IgG-Es (rS = 0.46, p = 0.002), and with IgG-Bp/IgG-Pg ratio (rS = 0.5, p = 0.0009) in cases of malformations, but not in women with normal pregnancy. Conclusion: we have revealed novel associations between congenital malformations of fetus and ratios of IgA-Bp/IgA-Es, as well as IgA-Bp/IgA-Pg, like as positive correlations between hormonal Pg/Es proportions, and ratios of specific antibodies in pregnant women.

The aim of the study was to evaluate FoxP3+ regulatory T cell (Тreg) subsets in the follicular fluid (FF) of the women undergoing in vitro fertilization (IVF), and their relationships with features of folliculogenesis and oogenesis, as well as quality of embryos and retrospective assessment of IVF outcomes. The study included 53 women at the age of 25 to 46 years stimulated by superovulation. The count of Tregs in the FF samples was determined by flow cytometry using appropriate monoclonal antibodies. The FF examination revealed of FoxP3+ T cells from both CD4+ (CD4+FoxP3+) and CD4(CD4-FoxP3+) T lymphocyte subsets. Moreover, the FoxP3+ cells were registered in CD4+CD25+ and CD4+CD25T lymphocyte subsets. Follicular fluid of women with a relatively small number of follicles contained higher numbers of CD4+CD25-FoxP3+ T cells, whereas a reduced number of oocytes was associated with the highest count of CD4-FoxP3+ T cells in FF. Retrospective analysis showed the a relationship between percentage of Tregs, and quality of oocytes and embryos. High fertilization index (0.75-1.0), reflecting maturity of the oocytes was associated with higher count of CD4-FoxP3+ T cells in the FF. Better quality of blastocysts was associated with a higher count of both CD4-FoxP3+ and CD4+FoxP3+ T cells. Onset and progression of pregnancies was also registered in women with relatively high counts of CD4-FoxP3+ T cells. The obtained data showed presence of different FoxP3+ T cell subtypes in follicular fluid, and its possible role in controlling early stages of reproductive process. CD4-FoxP3+ T cells seem to be the most important subpopulation; their counts are associated with efficacy of oogenesis, blastulation and pregnancy occurence.

Chemotherapy is among the primary methods of treating advanced breast cancer. It was shown that clinical efficacy of various chemotherapeutic agents in many cases depends not only on their direct cytostatic and/or cytotoxic effect upon tumor cells, but also on their ability to modulate phenotype of the tumor cells and to influence anti-tumor immune response. Initial state of the immune system and its response to treatment is crucial. Antitumor response involves cells of innate and adaptive immunity (NK, NKT, T cells). These populations are heterogeneous and contain, e.g., cells with antitumor activity and regulatory (suppressor) cells that suppress immune response and promote tumor progression. The aim of this work was to determine the relationship between the initial state of cellular immunity of patients suffering from locally advanced breast cancer with triple negative phenotype, and clinical effect of chemotherapy (cisplatin + doxorubicin/paclitaxel), and studying effects of the therapy upon subpopulation profiles of peripheral blood lymphocytes in the patients. We registered the terms of the disease progression as well as overall survival and progression-free survival. The disease progressed in 25 of 53 cases (47.2%) whereas 28 of 53 patients (52.8%) remained progression-free. The observation period was 35.5 months. Laboratory examination of the patients included immunophenotyping of peripheral blood lymphocytes and determination of NK cell cytotoxic activity before and after chemotherapy. Percentages of effectors and regulatory lymphocyte populations were determined. The results obtained showed that, for some lymphocyte subsets, the pre-treatment differences of cell percentage deviations from control were found between the progression-free groups and patients with progression of the disease. The differences in percentages of NKT cells and lymphocytes expressing CD25 activation marker proved to be most significant. Decreased number of NKT cells and activated CD25+ lymphocytes prior to chemotherapy was associated with increased probability of disease progression. Reduced percentage of NKT cells against control was observed in 56% of patients from the progression group (PD), and only 21.4% in the group free of disease progression (DF). [OR = 4.6 (95% CI 1.4 to 15.4)]. Percentage of CD25+ lymphocytes was decreased from 68.2% in the PD group, and 28.6% for DF patients [OR = 5.4 (95% CI 1.6-18.1)]. We studied relationships between the overall survival (OS) and percentage of perforin-containing NK, NKT, and T cells, and mean perforin fluorescence density (PFD) in these lymphocyte subsets in 26 of the 53 patients before treatment. A statistically significant positive correlation was revealed between OS and perforin PFD in all the three cell populations under study. Normalization of the parameters altered before treatment, and an increase of T cell numbers was observed in the disease-free patients.

At present, only ductal carcinoma in situ is included into the group of precancerous lesions of mammary ducts, according to International Agency for the Study of Cancer. However, based on recent publications, in addition to ductal carcinoma in situ, sclerosing adenosis, intraductal proliferative lesions and radial scar may be also attributed to precancerous changes. A variety of both benign and malignant events in mammary gland, the features of neoplastic growth and age of the patients require new approaches to study of carcinogenic events in mammary gland. As based on the known role of cytokines in genesis of malignancies, the aim of the study was to evaluate the cytokine-producing resource of immunocompetent blood cells in malignant, benign and precancerous mammary disorders. To assess the cytokine-producing resource of immunocompetent blood cells in the patients, we studied quantitative effects of polyclonal activators upon production of cytokines by immunocompetent blood cells of patients with invasive ductal cancer representing a histological type of adenocarcinoma (group I), and patients with non-malignant breast neoplasias (group II). At subsequent step, the patients with non-malignant neoplasms of the breast were divided into a subgroup of patients with only fibroadenoma and mastopathy (group III), and a group which included patients with precancerous diseases, i.e., sclerosing adenosis and interductal proliferates (group IV). Concentrations of IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF, and MCP-1 were determined by solid-phase enzyme immunoassay. When comparing groups I and II, we revealed higher influence of polyclonal activators upon production of G-CSF and GM-CSF in patients with invasive ductal cancer. When comparing the influence of polyclonal activation for cytokine production in patients of I and III groups, higher values were registered in patients with invasive ductal cancer (production of IL-2, G-CSF, and GM-CSF), and in patients with fibroadenoma and mastopathy (IL-18, and TNFαproduction). When comparing patients of groups I and IV, higher indexes of the polyclonal activator effects were found only for IL-1ra, G-CSF, and VEGF production in invasive ductal cancer. When comparing the indexes of polyclonal activator influence upon cytokine production of groups III and IV, higher values were obtained in patients with benign changes for the following cytokines: IL-8, IL-18, IL-1β, IL-1ra and TNFα, in contrast to patients with sclerosing adenosis and proliferates. The lower indexes of polyclonal activating effects upon the production of a number of cytokines in patients with precancerous changes, as compared to patients with malignant and benign breast tumors, do not indicate a decreased functional activity of immunocompetent blood cells. However, those may be due to high level of spontaneous cytokine production in sclerosing adenosis and interductal proliferates.

Uterine leiomyomas are common benign tumors developing from smooth muscle tissues, often leading to infertility and recurrent abortions. Pregnancy and development of uterine fibroids are characterized by an unusual rate of myometrium growth, hyperproduction of extracellular matrix and increased expression of numerous growth factor receptors. The purpose of this study was to determine concentrations of some key growth factors (IL-5, IL-7, IL-9, FGF-β, G-CSF, VEGF and PDGF) in blood serum of women with uterine myoma. Concentrations of the 27 cytokines were determined using a Bio Rad kit (USA) – Bio-Plex Pro™. Human Cytokine 27-plex Assay by means of flow-through fluorometry at the Bio-Plex 200 double-beam laser analyzer. Thirty-six patients with verified uterine myoma were followed up, being later subject to operative treatment (laparoscopic myomectomy). The results of this study showed a trend to decreased amounts of some hematopoiesis and angiogenesis growth factors, e.g., IL-9 and FGF, in blood serum of women with uterine myoma. Сoncentrations of IL-5, IL-7, and G-CSF proved to be significantly decreased if compared to serum contents of European healthy women. The most significant decrease was registered for pro-angiogenic factors, such as VEGF and PDGF. Their serum concentration in women with leiomyoma was reduced, respectively, 3- and 6-fold against controls. The decreased G-CSF concentration was not only quite significant, as compared to healthy women, but showed significant correlations with changes of such factors as IL-5, IL-7 and IL-9, with correlation quotients of, resp., 0.723, 0.637, and 0.504, respectively. One may conclude that, in spite of literature data on significantly increased contents of the mentioned growth factors in tissues of myometrium and growing uterine leiomyoma, our data show that the concentrations of these regulatory proteins in blood serum are decreased to some extent in this clinical condition.

Myeloperoxidase is a key factor promoting development of halogenative/oxidative stress under inflammatory conditions. Previously, we have discovered complexes including myeloperoxidase and its physiological inhibitor, ceruloplasmin in blood plasma of patients with inflammatory diseases of different etiology, e.g., atherosclerosis. Studies on regulation of myeloperoxidase activity by ceruloplasmin have shown that hypochlorous acid, a specific product of myeloperoxidase action, is likely to modify ceruloplasmin during inflammation. The present study was aimed for analysis of relationships between the myeloperoxidase activity, native, and HOCl-modified ceruloplasmin levels in blood plasma samples of the patients with cardiovascular diseases.

Specific antibodies against myeloperoxidase, ceruloplasmin, and HOCl-modified ceruloplasmin were obtained and specific enzyme-linked immunosorbent assays (ELISA) were developed. A combination of highly sensitive methods of myeloperoxidase assay i.e., solid-phase adsorption of antigens with subsequent testing of either their activity, or peroxidase-labeled antibody activity allowed elaborating the highly sensitive assays for ceruloplasmin and its HOCl-modified molecules, and for myeloperoxidase (concentration, peroxidase and halogenating activity). Positive correlation was proven between the myeloperoxidase concentration and activities. HOCl-modified ceruloplasmin content also correlated with myeloperoxidase activity.

The HOCl-modified ceruloplasmin was first discovered in blood plasma samples from patients with cardiovascular diseases. In view of correlation between myeloperoxidase activity and HOCl-modified ceruloplasmin content in plasma, we suggest that HOCl production is aimed for suppression of myeloperoxidaseinhibitory function of ceruloplasmin.

Insufficiency of local immunity can play an important role in pathogenesis of sepsis, including septic (acute) infectious endocarditis (IE). The paper presents data on secretory immunoglobulin A (sIgA) contents in blood serum of patients with sepsis (26 women and 32 men), acute (11 women and 23 men) and subacute (7 women and 13 men) IE, depending on localization of the infection site (angiogenic or non-angiogenic), outcome of the disease and carriage of glutathione-S-transferase P1 gene variants (GSTP1Ile105Val). A control group consisted of 25 women and 24 men without hypertension and ischemic heart disease and lacking evidence of focal and systemic infection, was examined. Laboratory studies were performed with еnzyme immunoassay and allele-specific polymerase chain reaction. We have found that, despite large individual variability of serum sIgA concentration in sepsis and infectious endocarditis, the majority of patients had a significant (on average, 4-fold) IgA increase against controls, in both men and women, especially in acute IE (a mean of 5-fold over control values). Subacute infectious endocarditis is associated with lesser sIgA in circulation than acute IE and sepsis, which may be used for early differential diagnosis of these conditions. There were no gender differences in sIgA contents. In sepsis with non-angiogenic source of infection, the sIgA levels were higher than in angiogenic infection. There was no association of sIgA level with survival (mortality), which excludes this index from predictive markers in sepsis and IE. Carriage of heterozygous GSTP1Ile105Val genotype increases the risk of sepsis and IE development, regardless of clinical course, and homozygous genotype GSTP1Ile105Ile is associated with higher contents of circulating immunoglobulin than in carriers of GSTP1Val105Val genotype. Thus, a wide range of individual variability in of circulating sIgA levels in patients with sepsis and infective endocarditis may be connected with location of infection source and genetic factors.

(IBD), who were for the first time treated with TNFα blocker (infliximab). Our aim was to determine prognostic informative value of the immunological parameters in order to assess the treatment efficiency. A comprehensive research included seventy children with IBD from 12 to 18 years old in the course of specific treatment (49 children with CD, 21 children with UC).

The comparison group consisted of fifty healthy children of similar age who were subjected to a similar detailed examination. The patients were divided into two groups, depending on their therapeutic response following 1 year of biological therapy: the first group showed a persistent positive effect of the drug, and the second group exhibited only unstable effects of the treatment. We determined the contents of major and small subpopulations of peripheral blood lymphocytes before the first administration of infliximab. Immunophenotyping was performed by multicolor flow cytometry (FC 500), using the CD45, CD3, CD4, CD8, CD19, CD16, CD56, HLA-DR, CD5, CD161, CD127, CD25, and CD294 markers.

We have revealed that the content of B lymphocytes was significantly reduced in children with unstable effects of therapy. By contrast, the B lymphocyte levels in children with persistent positive therapeutic effect did not differ from the comparison group. Analysis of the composition of the B lymphocyte profile showed an imbalance in the B1-to-B2 cell ratio, with decreased of B1 cell counts in IBD patients against the comparison group. In addition, the patients with unstable therapeutic effect showed a significant decrease in B2 cell numbers compared with a group with persistent effect and comparison group. The numbers of NK cells in IBD patients were found to be reduced against the comparison group. Assessment of T lymphocytes subsets revealed a number of features in the patients with minimal therapeutic effects, i.e., an increased level of activated T helper cells (CD4+CD25+CD127high) and Th17 lymphocytes (CD3+CD4+CD161+), as compared to children with stable effect of treatment and to the comparison group. Moreover, in children with minimal effects of therapy, the levels of Tregs within T-helper cell subsets were significantly higher than in the comparison group. By means of ROC analysis, we have identified most informative parameters for the groups with minimal versus persistent therapeutic effect, and showed a good quality for a discrimination model involving relative amount of Th17 cells, activated T helper cells and B lymphocytes. The number of Тh17 lymphocytes (% CD3+CD4+ lymphocytes) allowed to predict the effect of therapy with a TNFα blocker with high probability. The present study enables us to propose cellular immunity testing, as a promising tool for monitoring clinical state of IBD patients.

Pathogenetic mechanisms of uterine leiomyoma, adenomyosis and their combination are complicated and poorly understood, a differential diagnosis of leiomyosarcoma of the uterus is difficult. Our study aimed for a comparative analysis of the serum contents of α2-MG, PAG, some cytokines, sex steroids and the expression of steroid receptor genes in the patients with different variants of uterine proliferative diseases, in order to determine their pathological role, diagnostic and prognostic value.
Expression of estrogen receptor genes adenomyosis nodes was 1.5 to 2-fold higher than in leiomyoma, the combined pathology showed intermediate values, and expression of ER and PGR genes in leiomyosarcoma was minimal. In cellular leiomyoma, expression of ER receptor genes in the surrounding myometrium was 2 to 3-fold higher than in cases of simple leiomyomas. At the same time, concentration of estrogen and progesterone in the blood is comparable between the groups and control groups. All the patients have a deficiency of immunomodulatory α2-MG (12-13% for leiomyomas, 20% for adenomyosis, and 23% for malignant pathology). The concentration of immunosuppressive PAG is increased in combined conditions and leiomyosarcoma. In addition, the contents of IL-6 and TNFα increase, the VEGF levels exceed normal values 4 to 4.5-fold, in leiomyoma, 5.5-fold, in combined pathology, 6.5, in adenomyosis, and 10-fold, in leiomyosarcoma.
The obtained results confirm that immunomodulatory proteins, cytokines and cell-targeting sex hormones exert an interdependent influence upon each other in the studied diseases, and their significant changes may be used in diagnostics and prognosis.

In the present study, we have investigated frequency of genotypes and functional alleles of genes encoding chemokines (CXCL12 rs1801157, CCL2 rs1024611), chemokine receptors (CCR5 del32, CX3CR1 rs3732378), acute phase proteins SAA rs1136743, and CD14 rs2569190 polymorphisms among Tatar obese or overweight women from the Republic of Bashkortostan.

The group of patients comprised unrelated women with obesity (BMI ≥ 30 kg/m2, n = 225), females with overweight (BMI 25.0-29.9 kg/m2, n = 184), and control group of women (n = 327) BMI < 25.0 kg/m2. Genotyping was performed by PCR-RFLP analysis. Patients and controls differed in such parameters as body weight (p = 0.00001), BMI level (p = 0.001) and fasting glucose level (p = 0.0001).

An association was revealed between obesity and AG-AA genotypes (p = 0.007) and A allele (p = 0.003) of polymorphic locus rs3732378 of CXCR1 gene, as well as TT genotype (p = 0.027) and T allele (p = 0.021) of polymorphic locus rs1136743 of SAA gene. It has been shown that the AA genotype of polymorphic locus rs3732378 of the CX3CR1 gene is associated with increased body weight (p = 0.002) and elevated BMI (p = 0.018); the GG genotype of polymorphic locus rs1024611 of the CCL2 gene is associated with elevated fasting glucose level (p = 0.001).

As based on clinical and genetic data and using logistic regression, some statistically significant differences were revealed, which allow to predict development of obesity in Tatar women.

Granulysin and cathelicidin, the cytolytic molecules of innate immune system are important protective factors during infection with Mycobacterium tuberculosis. We present original data concerning high levels of granulysin and cathelicidin among the group of children and adolescents with latent TB infection. Patients with tuberculosis of the respiratory system exhibit significantly lower amounts of serum cathelicidin in destructive forms, as well as granulysin levels in “minor” forms of tuberculosis before starting the specific chemotherapy. The chemotherapy performed did not influence the serum granulysin and cathelicidin contents in patients with destructive tuberculosis, whereas the patients with “minor” forms (TLN/focal tuberculosis) revealed a significant increase in granulysin content after 6 months of treatment, and same trend for cathelicidin concentrations after 3 months of chemotherapy, followed by subsequent return to baseline values.

Laboratory diagnosis of antiphospholipid syndrome (APS) is based on detection of antiphospholipid antibodies (aPLs). E.g., aPLs are directed against conformational epitopes of the so-called “co-factor” proteins: β2-gycoprotein 1 (β2-GP1), annexin V (An V) and prothrombin (Pt) that are formed during interaction with phospholipids – cardiolipin (CL), phosphatic acid (Pha), phosphatidylcholine (Pch), phosphatidylethanolamine (Pe), phosphatidylglycerol (Pg), phosphatidylinositol (Pi), phosphatidylserine (Ps). A routine methodology of detection based on ELISA testing is challenged by new tests when the antigen is absorbed on another kind of support like microbeads or membranes that can influence density of conformational epitopes for aPL’s binding. The aim of our study was to compare the results of aPLs detection by ELISA and multi-line immunodot assay (MLD).
We collected blood serum samples from 45 patients with noncardioembolic ischemic strokes, 19 patients with recurrent deep vein thrombosis of lower limbs, 44 females with recurrent miscarriages, and 50 clinically healthy donors. To compare the results of aPL detection by ELISA and MLD kits, the test systems from different manufacturers were evaluated. We used an ELISA kits for detection of antibodies to CL IgG, aCL IgM, β2-GP1 produced by Euroimmun AG (Mr1) and Orgentec Diagnostica GmbH (Mr2) and MLD – for detection of antibodies to CL, β2-GP1, Pch, Pe, Pg, Pi, Ps, AnV and Pt (Medipan GmbH, Mr3).
When a cut-off titer was used as the main index, 30.5% of patients were aPLs-positive with ELISA method by Mr1 and 38%, wiht Mr2. By MLD aPls were detected in 30% of patients. In the same cohort, medium and high aPLs titers (> 40 U/mL) were determined in 12% of patients using ELISA kits. Positive and highly positive aPLs titers were determined in 16% when using a new method by Mr3. Medium and high titer were detected only for antibodies to β2-GP1, CL, An V, Pha and Phs.
The use of ELISA approach for detection of aPLs in patients with thrombosis and obstetric pathology is associated with relatively high number of low-positive ELISA results. Due to higher sensitivity for medium and high aPLs titers, MLD testing may be used as a confirming method for APS diagnosis.

Cytokines are the most important factors in pathogenesis of infectious, allergic, autoimmune, lymphoproliferative diseases and immunopathological processes. Many cytokines are very useful therapeutic targets for immunodiagnostics of different human diseases. Measurement of the cytokine levels by immunochemical methods in various biological fluids is usually used for diagnostic evaluation.
Content analysis of research articles from two Russian immunological journals, “Meditsinskaya Immunologiya” = “Medical Immunology (Russia)” and “Infektsiya i immunitet” = “Russian Journal of Infection and Immunity,”, shows that ELISA, xMAP multiplex immunoassay, and CBA technologies are the most common methods used in clinical and immunological studies aimed for determination of cytokine contents in blood serum/plasma.
Normal ranges of some plasma/serum cytokines in healthy individuals were subject to wide variations when using different methods and specific reagents from various manufacturers. The normal ranges applied by the CBA-technology, are significantly higher than appropriate values obtained by ELISA or xMAP-technologies. Most studies included a small control group, usually limited by 15-20 persons. In most of these works, blood serum samples were used for assays, whereas EDTA-conserved plasma was taken only in few studies.
It has been concluded that the results of cytokine measurements in blood serum/plasma in healthy individuals vary in wide ranges, and depend on many factors, e.g., initial sampling material, mode of technology, type of test systems, and characteristics of the group under study: number of patients, age, gender, geographical factor, etc. The mentioned data demonstrate a need for large-scale multicenter clinical studies, in order to standardize measurements of the cytokine levels in human peripheral blood and to specify their normal values.