Plasma cytokine profile in synucleinophaties with dementia.

Immune response may play a pivotal role in the pathogenesis of the common synucleinopathy as Parkinson's disease (PD) and could be mediated with the accumulation of neurotoxic alpha-synuclein. There is limited evidence for immune response in another synucleinopathy as dementia with Lewy bodies (DLB). Recent data suggest that immune response may contribute to cognitive impairment. We aimed to estimate plasma cytokine profile in patients with synucleinopathies with dementia (PD dementia (PDD), DLB). Plasma cytokine levels (interferon-gamma (IFN-gamma), interleukin (IL)-4 (IL-4), IL-6, IL-10, tumor necrosis factor alpha (TNF-alpha), monocyte chemoattractant protein-1 (MCP-1)). were estimated in 16 patients with DLB, 19 patients with PDD, 28 patients with PD without dementia (PD) and 19 individuals without neurological disorders (controls) using Luminex array system. Cognitive status was assessed with the Mini-Mental State Examination (MMSE). TNF-alpha and IL-6 plasma levels were elevated in patients with synucleinopathies with dementia (DLB, PDD) compared to controls and IL-10 plasma level was increased in PDD compared to controls (p < 0.05). IFN-gamma levels were decreased in PD and PDD patients compared to controls (p < 0.001, p = 0.026, respectively) and in PD patients than in DLB patients (p = 0.032). Patients with PD, PDD, and DLB were characterized by increased plasma levels of MCP-1 compared to controls (p < 0.001). At the same time, no differences in TNF-alpha, IL-10, IL-6 plasma levels in PD patients compared to controls were found. Our study demonstrated more pronounced immune response in synucleinopathies associated with dementia compared to PD without demetia.

Plasma Cytokines Profile in Patients with Parkinson's Disease Associated with Mutations in GBA Gene.

Plasma cytokine concentration in patients with Parkinson's disease and mutation in GBA gene, in patients with sporadic Parkinson's disease, and in healthy volunteers were measured by ELISA and multiplex analysis. In patients with Parkinson's disease and mutation in GBA gene, elevated plasma concentrations of IL-1ß and TNFα were revealed by ELISA in comparison with both controls and patients with sporadic form of Parkinson's disease. Multiplex analysis revealed enhanced secretion of IL-1ß, IL-2, IFNγ and reduced plasma levels of monocyte chemoattractant protein-1 (MCP-1) in patients with Parkinson's disease and mutation in GBA gene (in comparison with other groups) and increased plasma levels of IL-13 (only in comparison with the healthy volunteers). Our results support the hypothesis that the concentrations of inflammatory mediators are increased in patients with Parkinson's disease and mutation in GBA gene.

[THE STANDARD VALUES OF SUB-POPULATIONS OF T-HELPERS OF DIFFERENT LEVEL OF DIFFERENTIATION IN PERIPHERAL BLOOD].

The study was carried out to develop standard indicators of relative and absolute content of main populations of T-helpers in peripheral blood of conditionally healthy donors. The examination was implemented to sampling of 52 healthy individuals (29 males and 23 females) aged 18-65 years (median is 30 years). The multicolor cytofluorimetric analysis was applied using panel of following antibodies: CD45RA-FITC, CD62L-PE, CCR4-PerCP/Cy5.5; CCR6-PE/Cy7, CXCR3-APC, CD3-APC-AF750, CD4-Pacific Blue and CXCR5-Brilliant Violet 510TM. The T-helpers 1 were distributed in populations of cells with phenotypes CXCR5-CXCR3+CCR6-CCR4-, also containing Th9, and CXCR5-CXCR3+CCR6+CCR4- referred as Thl/Thl7. The Th2 were detected an the basis of availability of CCR4 at the absence of all other chemokin receptors. The Thi7, besides Thl/Thi7 mentioned above, were detected in composition of CXCR5-CXCR3-CCR6+CCR4- and CXCR5-CXCR3-CCR6+CCR4+. The last population also contained Th22. The follicular Th which expressed at their surface CXCR5, formed six cellular populations with following phenotypes: CXCR5+CXCR3-CCR6-CCR4- (Tfh/Tfh2), CXCR5+CXCR3-CCR6-CCR4+ (Tfh2), CXCR5+CXCR3-CCR6+CCR4- (Tfh17), CXCR5+CXCR3-CCR6+CCR4+ (Tfh17), CXCR5+CXCR3+CCR6-CCR4- (Tfh1) and CXCR5+CXCR3+CCR6+CCR4- (Tfh1/Tfh17). The relative and absolute content of T-helpers of mentioned phenotypes was established both within the framework of total population CD3+CD4+ of lymphocytes and among "naive" T-helpers (CD45RA-CD62L+), T-helpers of central (CD45RA-CD62L+) and effector (CD45RA- CD62L-) memory and also "terminal-differentiated" CD45RA-positive cells of effector memory with phenotype CD45RA+CD62L-. The study results can be applied as standard indicators under diagnostic of pathologic conditions of immune system.

Analysis of Populations of Memory T-Helper Cells Expressing CXCR3 and CCR6 Chemokine Receptors in Peripheral Blood of Patients with Chronic Viral Hepatitis C.

Flow cytometry was employed to examine the content of major populations of memory T helper cells and expression of chemokine receptors CXCR3 and CCR6 on their surface in peripheral blood drawn from virtually healthy people and the patients with chronic viral hepatitis C. The following combination of monoclonal antibodies had been used: CD62LFITC/CD45RA-PE/CD3-ECD/CCR6-PC7/CXCR3-APC/CD4-APC-Cy7. In comparison with control group, the patients with chronic hepatitis C had a smaller number of populations of naïve CD4(+) T cells and central memory CD4(+) T cells but a greater number of terminally differentiated effector memory CD4(+) T cells and effector memory CD4(+) T cells. No differences were revealed between CD4(+) T cell populations of both groups in expression of CXCR3 and CCR6 receptors.

Chemokine Receptors CXCR3 and CCR6 and Their Ligands in the Liver and Blood of Patients with Chronic Hepatitis C.

We performed a comprehensive analysis of CCR6 and CXCR3 chemokine receptors and their ligands CCL20/MIP-3α, CXCL9/MIG, CXCL10/IP-10, and CXCL11/ITAC in the liver and blood of patients with chronic hepatitis C at different stages of the disease. TaqMan PCR was used to determine mRNA gene expression of chemokines and their receptors in liver specimens, xMAP multiplex analysis was performed to estimate the concentration of chemokines in blood plasma, and fl ow cytofluorometry was used to evaluate CCR6 and CXCR3 expression on peripheral blood lymphocyte populations. In the liver of patients with hepatitis C, mRNA expression of CXCL10, CCR6, and CXCR3 genes increases with fibrosis progression in the liver tissue. In the plasma, concentrations of all studied chemokines increased depending on the stage of liver fibrosis, CCR6 and CXCR3 expression was changed in various lymphocyte populations. Thus, chemokines are involved in the immunopathogenesis and fibrogenesis in chronic viral hepatitis C. The results suggest using these chemokines in the diagnosis and prognosis of the disease.

[THE ROLE OF CYTOKINES AND CHEMOKINES IN LABORATORY DIAGNOSTIC OF CHRONIC VIRAL HEPATITIS C].

The chronic viral hepatitis C is widely prevalent disease with prolonged persistence of virus and obliterated clinical picture. The present techniques of diagnostic of degree of fibrosis of liver and prognosis of course of disease have particular shortcomings. Hence, search of safe low invasive methods based on blood biomarkers is an actual task. The cytokines/chemokines (mediators of chronic inflammation) directly involved into immunopathogenesis of chronic viral hepatitis C can act in the capacity of biomarkers. The study was carried out to comprehensively analyze content of cytokines/chemokines in peripheral blood of patients with chronic viral hepatitis C at various stages of disease and infected by different genotypes of virus of hepatitis C. The concentration of cytokines/chemokines was identified in blood plasma of patients with chronic viral hepatitis C (n = 73) and conditionally healthy donors (n =3 7): IFNα, IFNγ, IFNλ/IL28α, TNFα, CCL2/MCP-1, CCL3/MIP-lα, CCL4/MIP-lß, CCL5/RANTES, CCL8/MCP-2, CCL20/AIP-3α, CXCL9/MIG, CXCL10/P-10, CXCLII/ITAC. The multiplex analysis using technology xMAP was applied. The increasing of level of TNFα, CCL2/MCP-1, CCL4/ MIP-l, CCL8/ACP-2, CCL20/MIP-3α, CXCL9/MIG, CXCL10/IP-10, CXCL11/ITA C was established in blood plasma of patients with chronic viral hepatitis C as compared with control group. The levels of analyzed interferons IFNα, IFNγ, IFNλ/IL28α had no difference in studied groups. As far as chronic viral hepatitis C progresses and fibrosis of hepatic tissue develops the concentrations of TNFα, CCL2/MCP-1, CCL20/MIP-3α, CXCL9/MIG, CXCL10/IP-l0, CXCL11/ITAC increased significantly. The concentrations of chemokine CXCL11/IT4 C can be used as informative indicator for differentiating diagnostic of early stages of liver fibrosis. Depending on genotype of virus of hepatitis C, in patients with chronic viral hepatitis C change in content of CCL8/MCP-2 was established. Hence, detection in blood plasma of patients with chronic viral hepatitis C concentration of particular cytokines/chemokines using multiplex analysis technique permit analyzing additional information concerning degree of liver fibrosis, activity of process of damage of hepatic tissue under chronic viral hepatitis C that indicates indirectly on genotype of virus of hepatitis C.

[The comparative value of quantiferonic test, neopterin and specific anti-tuberculosis antibodies in clinical diagnostic of tuberculosis of lungs].

The article deals with comparison of clinical diagnostic value of laboratory techniques of tuberculosis. The sample included 63 patients with tuberculosis of lungs. 49 persons of long-time contact with patients with tuberculosis and 28 healthy donors. The QuantiFERON-TB Gold In-Tube test was applied to total sampling. The content of neopterin and specific anti-tuberculosis antibodies in blood plasma was determined. According study data, the mentioned test is mostly informative for detection of tuberculosis contamination (sensitivity 64%, specificity 89%). However, the obvious shortcoming of this technique is the fact that it can't provide the differentiation between active and latent tuberculosis infection. As opposed to QuantiFERON-TB Gold In-Tube test, the detection of specific anti-tuberculosis antibodies (sensitivity 54%, specificity 94%) and neopterin (sensitivity 51%, specificity 92%) makes it possible to differentiate these two groups of patients.