Transplantation of allogeneic hematopoietic stem cells is the only curative option for a number of diseases of hematopoietic system. It is intended to replace the hematopoietic system of the recipient with
the donor’s. However, when mature T cells contained in the graft enter the recipient organism, it may lead to a severe post-transplant complication, the “graft versus host” disease (GVHD). It occurs due to the fact that the donor immune system contains T cell clones specific to recipient alloantigens. These cell clones are activated upon encountering their antigens, thus causing systemic damage to healthy tissues. Development of the alloreactive clones is caused by genetic differences between donor and recipient. The most important
factors determining successful transplantation concern the compatibility for the genes coding for Major Histocompatibility Complex (MHC), that are expressed in all nucleated cells and are responsible for antigen presentation to the immune cells. Currently established extensive donor banks allow for majority of patients to choose a compatible donor. However, this does not provide complete prevention of the GVHD development, because in addition to the MHC genes the donor and recipient may differ in so-called minor histocompatibility antigens. Minor antigens may be caused by genetic polymorphisms in all of the genome coding regions. Pre-transplantation conditioning of the patient, which is necessary for engraftment represent an additional factor contributing to the GVHD
development, since as its side effect it leads to formation of a pro-inflammatory environment in the organism of recipient. Severe GVHD develops in approximately 40% of MHC-matched patients, while in cases of partial compatibility this proportion is even higher. GVHD causes mortality comparable to other causes of posttransplant
death, such as viral infections or relapse of underlying disease. Thus, the development of severe GVHD is a significant limitation for clinical applications of stem cell transplantation. Severe immune
suppression or depletion of mature donor T cells from the transplant leads to increased probability of relapse and weakens anti-infectious immunity. Hence, further search for alternative, more specific ways to prevent GVHD is required. This review will focus on the mechanisms of alloreactive T lymphocyte clone development and key pathogenetic stages of acute “graft versus host” disease.

Disorders of immune state in desynchronosis may be associated with reduced concentrations of melatonin in blood, thus being a prerequisite for pharmacological correction of appropriate homeostatic changes. The purpose of this work was to explore some mechanisms of exogenous melatonin actions upon parameters of innate and adaptive immunity in experimental model of desynchronosis under the conditions of LED illumination. The study was performed with 196 adult guinea pigs. Light desynchronosis was produced by day-and-night illumination of the animals having been continued for 30 days. Melatonin was administered applied per os daily at the total dose of 30 mg/kg. A solution of melatonin in isotonic NaCl solution was prepared from the Melaxen drug (INN: melatonin, “Unipharm Inc.,” USA) ex tempore. To study innate immunity of blood cells, we determined leukocyte numbers, WBC differential counts, and functional activity of phagocytes, as spontaneous and induced NBT test, as well as engulfment of polystyrene latex particles. Th1-specific immune response was studied according to degree of delayed type hypersensitivity reaction; Th2-dependent response was assessed as the numbers of antibody-forming cells in the spleen of the animals after immunization with allogeneic erythrocytes. Serum concentrations of interleukin 4 (IL-4), interferon-gamma (IFNγ), melatonin, and cortisol were measured by enzyme immunoassay, using the “Immulayt 2000” (USA) with guinea pigspecific test systems. It was found that experimental desynchronosis was associated with leukocytosis, lymphoand monocytopenia, activation of oxygen-dependent metabolism of blood phagocytes, suppression of Th1-and Th2-dependent immune response. Desynchronosis was also accompanied by decreased concentrations of serum melatonin, IFNγ and IL-4, along with increased cortisol concentrations. Reduced IFNγ and IL-4 amounts was associated with decreased melatonin concentrations. Suppression of Th1- and Th2-dependent immune response is found to develop in accordance with reduction in IFNγ and IL-4. Melatonin administration in desynchronosis (a total of 30 mg/kg) resulted in recovery of blood leukocyte counts, ROS generation by phagocytes, higher intensity of Th1- and Th2-dependent immune response, cortisol, IFNγ and IL-4 concentrations in blood serum, like as stimulation of blood phagocytic capacity. Restoration of Th1- and Th2-dependent immune response is associated with normalized concentrations of, respectively, IFNγ and IL-4 levels in blood serum.

Multiparameteric flow analysis has offered an ability of simultaneous analysis of multiple molecules at the single-cell level. Peripheral blood cells from 110 healthy subjects aged 18-65 years (59 males and 51 females) were stained with antibodies to CD3, CD4, CD8, CD27, CD28, CD45, CD45RA and CD62L, and analyzed using different gating strategies. The first one was based on initial analysis of CD45RA and CD62L expression, and CD3+CD8+ cells were divided into naïve population (CD45RA+CD62L+) comprising approx. 30% of the CD3+CD8+ subset; central memory cells (CD45RA–CD62L+, ~11%), effector memory cells (EM; CD45RA–CD62L–, ~35%) and «terminally differentiated» effector memory cells (TEMRA, CD45RA+CD62L–, ~24% of total CD8+ subset). As based on expression of CD27 and CD28 in EM and TEMRA, some further populations were distinguished, i.e., CD27+CD28+ (termed as EM1, about 19% from CD3+CD8+); CD27+CD28– (EM2, ~5%), CD27–CD28– (EM3, ~9%) and CD27–CD28+ (EM4, ~2%). Appropriate subsets were identified within TEMRA population, as follows: CD27+CD28+ (pE1, ~3%), CD27+CD28– (pE2, ~5%) and CD27–CD28– (E, ~15%). The second approach was based on initial
expression of CD27 and CD28, followed by analysis of CD45RA and CD62L expression on CD27+CD28+subset. Total cytotoxic T cell population was divided into naïve – CD27+CD28+CD45RA+CD62L+ (~30% from CD3+CD8+ subset), central memory (CD27+CD28+CD45RA–CD62L+, ca.~12% of total), transitional memory cells (CD27+CD28+CD45RA–CD62L–, approx.~12%), as well as effector memory cells and effector
cells (CD27+CD28я, ~11% и CD27–CD28–, ~24%, respectively). Expression of CD45RA and CD62L was not analyzed for the latter two populations. Frequencies of all cell populations, identified by means of two different gating strategies, were expressed as percentages of the total CD3+CD8+ and absolute cell counts. Using the gating strategy based on initial analysis of CD45RA and CD62L, some correlations between naïve CD3+CD8+frequencies and donor age were revealed (r = -0.646, р < 0.001, and r = -0.562, р < 0.001, respectively). Relative and absolute counts of ЕМ3 (r = 0.474, р < 0.001 and r = 0.435, р < 0.001, respectively) and Е subsets (r = 0.393, р < 0.001 and r = 0.375, р < 0.001, respectively) CD3+CD8+ subsets showed linear increase with age. Usage of another gating strategy based on CD27 and CD28 expression revealed age-dependent changes in relative and absolute frequencies of naïve CD3+CD8+ (r = -0.638, р < 0.001 and r = -0.530, р < 0.001, respectively). Meanwhile, CD27–CD28– subset accumulated linearly with age (r = 0.495, р < 0.001 and r = 0.442, р < 0.001, respectively). The results suggest that differences in subset distribution are responsible for age-related changes in CD8+ cells.

Cytokines play an important role in the pathogenesis of liver cirrhosis (LC), determining the disease severity as well as hepatic and extrahepatic complications. The present study aims to characterize the cytokine profile of blood cells in 20 LC patients with regard to the stage (class A on Child-Pugh, n = 13, and the class B+C, n = 7) and the etiology of the disease (viral LC, n = 12 and the LC of non-viral etiology, n = 8) using cytokine 26-plex assay. The control group consisted of 10 healthy donors. Cytokine concentrations were tested both in spontaneous and bacterial endotoxin (LPS) stimulated 24 hour blood cultures. Four functional groups: pro-/anti-inflammatory cytokines (IL-1β, TNFα, IL-1ra, IL-10), immunoregulatory cytokines (IL-2, IFNγ, IL-12, IL-4, IL-5, IL-6, IL-9, IL-13, IL-15, IL-17), growth factors (G-CSF, IL-7, FGF-β, PDGF, VEGF) and chemokines (IL-8, IP-10, MCP-1, MIP-1α, MIP-1β, RANTES, Eotaxin) involved in the pathogenesis of inflammation, have been analyzed. The blood cells of LC patients were shown to be active producers of cytokines and were characterized by increased cytokine levels from all four functional groups. The enhancement of many cytokines was associated with the disease severity and viral etiology. The increased cytokine production in both intact and LPS-stimulated cultures indicated preserved reactivity of blood cells to endotoxin. Overall, the data obtained suggest the contribution of circulating blood cells in supporting of inflammatory response and the potential diagnostic/prognostic significance of blood cell-derived cytokines in patients with LC.

Osteoarthritis (OA) is one of most common rheumatic diseases, and currently there is no effective pharmacological treatment of OA. It has been suggested that lack of effective treatment is, in part, due to the disease heterogeneity which may lead to development of several OA subtypes (phenotypes). Diabetes-associated OA is among the proposed OA phenotypes. The key mechanism involved into inflammatory and degenerative changes in OA is a decrease in DNA methylation suggested for several cell types, that was also demonstrated in type 2 diabetes mellitus. Therefore, pharmacological increase of DNA methylation may be an effective treatment strategy which may exert pleiotropic effects in diabetes-associated OA. In a randomized crossover study, we have evaluated efficacy and safety of ademetionine, a methyl group donor, in comparison with chondroitine sulfate in patients with OA associated with type 2 diabetes mellitus. The patients were randomly assigned to sequential treatment of chondroitine sulfate/ademetionine or ademetionine/chondroitine sulfate during one month, with a washout period of 2 weeks. The primary endpoint was pain measured according to visual analogue scale (VAS). Painful symptoms, as well as function and disease signs in knee, hip and hand joints were also assessed with KOOS, WOMAC, and FIHOA scales. General performance was assessed with SF–36 scale. To evaluate systemic inflammation, we measured serum IL-6, IL-18, adiponectin, and CRP using ELISA technique. Concentrations of serum cartilage destruction biomarkers (aggrecan and antibodies to collagen type II) were assessed by ELISA. Serum lipid levels were measured with standard method; glycated hemoglobin was assessed with liquid chromatography. Ten patients (all women, age 61.7-74.2 year with BMI of 1.1-38.4 kg/m2) were included in the study. It has been demonstrated that ademetionine showed a statistically significant analgetic effect (decrease in VAS pain), improved knee function and reduced symptoms in knee joints (as measured by KOOS subscales), and did not influence the levels of systemic inflammation or cartilage destruction biomarkers. There was also no change in lipid levels and glycated hemoglobin concentrations. Ademetionine was well tolerated, no serious adverse events occurred during the treatment. In conclusion, ademetionine does not have pleiotropic pharmacological effects in diabetes-associated OA. Its potential application in cases of different comorbidities requires further studies.

Early course of acute systemic inflammatory response after surgical intervention was traced for sixty-four patients with chronic calculous cholecystitis following a laparoscopic cholecystectomy (LCE). The patients were classified in 2 groups dependent on the surgery mode. The main group included 32 patients after single-port LCE, and controls that underwent four-port LCE (n = 32). Peripheral blood samples were taken 2 h before intervention, as well as 6, 24, and 48 h after surgical treatment. of Alpha-1-antitrypsin (A1AT), C-reactive protein, TNFα and IL-1β levels were measured in the specimens. For A1AT, we have not found any significant differences between the baseline and post-surgery levels, both in main and control groups. The intergroup comparisons showed a statistically significant increase in A1AT levels 24 h after the 4-port LCE. The 
patients subjected to single-port LCE, exhibited similar pre- and post-surgery levels of C-reactive protein. In the control group, its level proved to be statistically higher against initial values 6 h after the surgery. Comparing the both groups, C-reactive protein was found to be significantly increased in controls 6 и 24 h after surgery.
The post-op TNFα levels after the single-port LCE tended to increase, as compared to the baseline values. A statistical increase of TNF levels over initial values was noted 24 h after the 4-port LCE. Upon the intergroup comparisons, a significant TNF increase was revealed 24 following the 4-port LCE. IL-1β levels in the main group did not differ between the pre- and post-surgical period. However, its statistically significant increase to the initial values was revealed in controls 24 h after surgery. A comparison for IL-1β levels between the 2 groups has shown its significant elevation 24 h after the 4-port LCE. These data allow to conclude that a systemic inflammatory response as assessed by acute phase proteins in patients after a single-port LCE, is characterized by a shorter and less marked inflammatory reaction, as compared to the reaction in patients after the 4-port LCE.

Molecular basis still remains unclear for psoriasis, a chronic inflammatory skin disease. It biological features are presented by abnormal differentiation of epidermal keratinocytes, overgrowth and dilation of blood vessels, and leukocyte infiltration of dermal and epidermal skin layers. These events appear to be driven, mainly, by various cytokines and chemokines released by activated T cell populations. The aim of this replication study was to determine, whether the rs4649203 SNP of IL28RA gene is associated with susceptibility to psoriasis. A total of 341 patients with psoriasis and 407 matched healthy controls were enrolled to carry out a case control study. Genotyping was performed using a Real-Time PCR assay. Our preliminary data suggest that the polymorphism located in IL28RA gene, known to be related to inflammatory and immunity processes, showed an association with patients’ age at onset and the disease severity. The results of this study are promising, with respect to development of a personalized approach to psoriasis treatment.

Some basic parameters of immune profile have been investigated in 140 patients with chronic lymphocytic leukemia (CLL). There was shown that the cohort of CLL patients is heterogeneous for their immune profile as early as at primary diagnosis stage, showing sufficient differences in the numbers of immunocompetent cells in peripheral blood. In most cases, the interpretation of appropriate quantitative characteristics is quite difficult, if based on the conventional criteria. The numbers of activated T-lymphocytes and NK-cells were proven to be the most informative parameters of cellular immunity in this cohort. Progression of leukemic events shows a close association with only a single parameter under study, i.e., CD4+/CD8+ cell ratio.

The aim of present work was to study some activation features of neutrophilic granulocytes from peripheral blood of the patients with G type of multiple myeloma (MM), as depending on clinical stage of the disorder. In this study, a number of parameters were evaluated, as follows: time dynamics of the up-to-maximum luminescence increase; maximal luminescence levels, areas under the curves (AUC) for the dynamic luminescence registration. Zymosan was used as chemiluminescence inducer, and luminol, as enhancer of the “respiratory burst”. The chemiluminescence enhancement was assessed as a ratio of AUC values corresponding to induced-to-spontaneous chemiluminescence being denoted as an ‘activation index’. The onset and progression of MM in humans was accompanied by disturbance of neutrophil functions. MM patients at stage II and III exhibited a significant increase of spontaneous ROS production relative to controls. The inducible ROS production by neutrophils was also increased in all groups, as compared with spontaneous production and and control parameters, thus arguing for a distinct role of neutrophils, as potentially cytotoxic effectors against tumor cells, along with their basic function of nonspecific antimicrobial protection.