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Objective To explore the immunopathological mechanism for the imbalance between the positive signal mediated by inducible costimulator (ICOS) and the negative signal mediated by programmed death-1 (PD-1) in patients with myasthenia gravis (MG).Methods Eighty-two patients with MG,56 healthy controls (HC) and 20 non-MG (NMG) patients,collected in the First Affiliated Hospital of Suzhou University from February 2014 to December 2016,were chosen to participate in the study.The expression of ICOS and PD-1 on peripheral blood mononuclear cells was detected by immuno-fluorescence staining and flow cytometry.The levels of soluble programmed death-1 (sPD-1),soluble programmed death ligand 1 (sPD-L1),IL-4 and other cytokines were detected by enzyme-linked immunosorbent assay.Results (1) Flow cytometry analysis:The co-expression of PD-1,ICOS on CD4 + T cells from MG group (9.64％ (8.82％)) was higher than in HC (1.81％ (2.10％),Z =-7.389,P ＜0.05) and NMG group (2.86％ (1.49％),Z =-4.636,P ＜ 0.05).The expression of ICOS on CD4 + T cells,ICOS ligand (ICOSL) on CD14+ monocytes and CD19+ B cells were increased in MG group comparing with that of the control groups.The proportion of PD-1 + CD4 + T cells (MG group 16.82％ (10.66％),HC 9.34％ (9.18％),Z =-4.345,P＜0.05;NMG group 7.07％ (3.40％),Z=-4.594,P＜0.05) and PD-1 Ligand (PD-L1) + CD14+ monocytes was higher in MG patients.All of these were detected by flow cytometry.(2) ELISA analysis:Serum sPD-1 expression significantly increased in MG group compared with that in the control groups (MG group (1.87 ± 0.64) ng/ml,NMG group (1.49 ± 0.70) ng/ml,t =2.04,P ＜ 0.05;HC (1.05 ± 0.50)ng/ml,t =2.08,P ＜ 0.05),while for serum sPD-L1,there was no significant difference between MG and control groups.(3) Serum cytokines detection:The expression of IL-4 was increased in MG patients (MG group (61.88 ±5.15) pg/ml,HC (32.03 ±1.84) pg/ml,t=2.50,P＜0.05;NMG group (42.62± 3.31) pg/ml,t =2.34,P ＜0.05),and there was a negative correlation between the expression of sPD-1 and the concentration of IL-4.Conclusions The increased expression of PD-1 + ICOS + CD4 + T cells suggested the subset involved in the pathological progress of MG.sPD-1 might disturb the ligation of PD-1 on T cells and PD-L1 on antigen presenting cells,while the ligation of ICOS and ICOSL passed positive signal,leading to over activity of the subsets and the progression of disease.
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Objective To explore the mechanism of sulfotanshinone sodium injection in treatment of patients with sudden deafness (SD).Methods Sixty patients with SD admitted to the Department of Otorhinolaryngology of Wuxi Traditional Chinese and Western Medicine Hospital from January to December 2016 were enrolled, and they were randomly divided into a study group and a control group (each 30 cases). The same basic treatment was given in the two groups, the patients in the study group were treated with sulfotanshinone sodium 40 mg intravenous (IV) drip, while the patients in the control group were treated with vinpocetine sodium chloride 20 mg IV drip, once a day for consecutive 14 days to complete a therapeutic course, and two courses were carried out in bothgroups. Before and after treatment, the changes of hearing threshold, indexes of hemorheology and immune function were compared between the patients in the two groups, and the clinical efficacy and adverse reactions in the two groups were observed.Results After treatment, the hearing threshold, hemorheology indexes, immune function index of CD8+ were significantly lower than those before treatment, while the CD3+, CD4+, CD4+/CD8+ ratio were significantly higher than those before treatment in the two groups, and the above changes of indexes were more obvious in the study group than those in the control group hearing [threshold (dB): 16.63±2.04 vs. 17.15±1.88, plasma viscosity (PV, mPa·s): 1.27±0.14 vs. 1.31±0.11, whole blood middle shearing viscosity (mPa·s): 4.77±0.33 vs. 4.95±0.28, whole blood high shearing viscosity (mPa·s): 3.86±0.25 vs. 4.00±0.31, erythrocyte aggregation index (EAI): 1.57±0.29 vs. 1.72±0.34, CD3+: 0.70±0.05 vs. 0.64±0.05, CD4+: 0.43±0.04 vs 0.37±0.03, CD8+: 0.32±0.04 vs. 0.34±0.03, CD4+/CD8+: 1.36±0.32 vs. 1.18±0.27]; the degree of whole blood low shearing viscosity (mPa·s: 6.72±0.80 vs. 7.01±1.13) and hematocrit (HCT: 0.38±0.04 vs. 0.40±0.03) decreasing weremore significant in the control group than those in the study group. The total effective rate was higher in study group than that in the control group [86.67% (26/30) vs. 83.33% (25/30)], but the difference between the two groups was not statistically significant (P > 0.05); the incidence of adverse reactions in the study group was markedly lower than that in the control group [3.33% (1/30) vs. 20.00% (6/30),P < 0.05].Conclusions Sulfot anshinone sodium injection can effectively enhance the SD patients' hearing, and improve their hemorheology indexes and immune function; the therapeutic results of sulfotanshinone sodium injection in safety and improvement in immune function are superior to those of vinpocetine sodium chloride injection.
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Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.
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Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.
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Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.
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In this paper, the aging of the stem cells has been elucidated in the aspect of quantitative and qualitative exhaustion, such as balance self-renewal and differentiation of hematopoietic stem cells. The reduced telomere and oxidative stress both accelerate the aging course, besides, histone code and transcriptional activation lead to the DNA or protein damage. It plays an important role in the stem cell aging that the modulation of allochromosome structure and epigenetic regulation of gene transcription. If the aging of the stem cells doesn’t happen on the level of the gene, it must result from the transcription. During the haemopoiesis process of stem cells, all kinds of the transcriptional mistakes will lead to the aging of cell cycle, which is the result of allochromosome enlarging. However, the above mechanism will not occur normally. Only allochromosome allotopia is not explainable to the gene expression of stem cells. The proof of stem cells’ self-renewal evidences that if not the consequence of cell expression, allochromosome change the histone.