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Objective: To investigate the role and significance of immunoassay locus LAG3 gene in Alzheimer's disease (AD). Methods: Based on the GEO (https://www. ncbi. nlm. nih. gov/geo/) in the database of AD patients, brain sequencing dataset GSE48350 and blood sequencing dataset GSE140829 were analyzed. The scores of immune cells in the brain tissue were obtained by using the CIBERSORT ratings and the expression of LAG3 was observed. Mouse hippocampal neuron cell line HT22 induced by human Aβ1-42 amyloid was used as the cell model of AD disease. The expression of LAG3 was detected by RT-PCR. GSEA analysis was used to explore pathways directly associated with LAG3 and the String site was used to build PPI networks, genes interacting with LAG3 were discovered, and the biological functions of these genes were analyzed by KEGG and GO. Results: There was a great difference between the constitute of immune cells in the brain tissue of AD patients and that of normal controls. The level of LAG3 expression in brain tissue and blood of AD patients were higher than that of normal controls, so as the cell model. Conclusion: The constitute of immune cells in AD disease brain tissue and normal person is different. The results in the brain tissue of AD patients of the higher percentage of inactive memory CD4+T cells and mononuclear cells, lower proportion of the activation of NK cells, and the more increased expression of immune examination site LAG3 may be associated with CD4 cells and the activation and regulation of immune cells. [ABSTRACT FROM AUTHOR]

Objective To evaluate the changes and significance of lymphocyte subsets in the recipients with acute rejection after liver transplantation. Methods The recipients presenting with acute rejection after liver transplantation were assigned into the rejection group (n=17), and their counterparts with stable liver function were allocated into the control group (n=17) according to the ratio of 1∶1 by propensity score matching method. The incidence of acute rejection after liver transplantation was analyzed, and the concentration of tacrolimus in the recipients was compared between two groups. The absolute value and proportion of lymphocyte subsets in peripheral blood were compared between two groups. The diagnostic value of lymphocyte subsets for acute rejection after liver transplantation was assessed by the receiver operating characteristic (ROC) curve. The absolute value and proportion of lymphocyte subsets in the rejection group were compared before and after treatment. Results Among 17 recipients in the rejection group, 4 cases developed acute rejection within postoperative 28 d, and 13 cases had acute rejection within postoperative 29-180 d. No significant difference was noted in the tacrolimus concentration between two groups (P=0.295). Compared with the control group, the proportions of peripheral blood T cells, CD4+ T cells, B cells and natural killer (NK) T cells were significantly increased in the rejection group (all P<0.05). The elevated proportion of NKT cells in the early stage after liver transplantation was an independent risk factor for acute rejection following liver transplantation[odds ratio (OR) 1.774，95% confidence interval (CI) 1.059-2.971，P=0.029]. ROC curve analysis showed that the area under curve (AUC) of CD4+ T cells, B cells and NKT cells was 0.76, 0.73 and 0.77, respectively. The AUC of combined use of CD4+ T cells, B cells and NKT cells was 0.89, with a cut-off value of 0.69, sensitivity of 0.706 and specificity of 0.941. After corresponding treatment, all recipients were gradually recovered, and liver functions were eventually restored to normal in the rejection group. After treatment, the proportion of T cells, CD4+ T cells, CD8+ T cells and NK cells was significantly decreased (all P<0.05). Conclusions The elevated proportion of NKT cells indicates an increased risk of acute rejection after liver transplantation. Combined use of CD4+ T cells, B cells and NKT cells may deliver early detection and diagnosis of acute rejection after liver transplantation. [ABSTRACT FROM AUTHOR]

Objective To investigate the role of CD4+ CD45RClow regulatory T cell (Treg) in the immune tolerance induction of rats undergoing liver transplantation. Methods Liver transplantation rat models of acute rejection (AR) [Lewis → Brown Norway (BN), AR group] and spontaneous tolerance (BN → Lewis, tolerance group) were established, with 6 rats in each group. Moreover, 3 Lewis rats and 3 BN rats were assigned into the sham operation group (control group). The liver tissues of rats in each group were subject to pathological staining. The expression of T cell subsets and plasmacytoid dendritic cells (pDC) in the peripheral blood, liver graft and spleen of rats was detected in each group. The correlation between pDC and CD4+ CD45RClowTreg was analyzed. The expression levels of CD4, CD45RC and CD103 in the liver graft and spleen of rats were quantitatively measured in each group. Results In the AR group, pathological manifestations mainly consisted of inflammatory cell infiltration and structure disorders of transplant liver. Compared with the AR group, the expression levels of CD4+ CD25+ Treg and CD8+ Treg in the peripheral blood were significantly upregulated in the tolerance group (all P<0.05). In the peripheral blood, the expression level of CD4+ CD25+ Treg was positively correlated with that of CD8+ Treg (r=0.742, P=0.022). In the AR group, the expression level of CD4+ CD45RChighT cell in the peripheral blood was significantly higher than those in the tolerance and control groups (both P<0.05). Compared with the AR group, the expression level of CD4+ CD45RClowTreg in the spleen, and the expression levels of CD8+ CD45RClowTreg in the peripheral blood, transplant liver and spleen were significantly up-regulated in the tolerance group (all P<0.05). Compared with the control and AR groups, the ratio of CD8+ CD45RClowTreg/CD8+ T in the peripheral blood and the expression levels of pDC in the peripheral blood, transplant liver and spleen were all significantly up-regulated in the tolerance group (all P<0.05). The expression level of CD4+ CD45RClowTreg was positively correlated with the changes of pDC (r=0.506, P=0.016). The expression levels of CD4, CD45RC and CD103 in the transplant liver and spleen of rats were up-regulated in the tolerance group. In the AR group, the expression levels of CD4 and CD45RC were up-regulated, whereas that of CD103 was down-regulated. Conclusions CD4+ CD45RClowTreg is a cell subgroup with negative immune regulation, which may construct a regulatory cell network of immune tolerance induction along with CD8+ CD45RClowTreg and pDC, [ABSTRACT FROM AUTHOR]

Objective To analyze the dynamic changes and the influencing factors of T lymphocyte subsets in recipients with stable graft status within 1 year after lung transplantation. Methods Clinical data of 41 recipients with stable graft status after allogeneic lung transplantation were analyzed. The absolute value and ratio of T lymphocyte subsets in peripheral blood from recipients were measured by flow cytometry before operation, 2 weeks and each month (within 1 year) after operation, respectively. The effects of age, gender, body mass index (BMI), surgical method, incidence of primary graft dysfunction (PGD) after operation, and primary disease upon the absolute values of T lymphocytes were evaluated. Results Within 1 year after lung transplantation, the absolute values of CD3+, CD3+ CD4+, CD3+ CD8+ T lymphocytes and CD4+ /CD8+ ratio were changed over time (all P<0.001). Compared with preoperative values, there was no statistical significance in the absolute values of CD3+ and CD3+ CD4+ T lymphocytes at 12 months after operation (P=0.659, 0.109), whereas the absolute value of CD3+ CD8+ T lymphocytes was increased (P=0.02) and the CD4+ /CD8+ ratio was decreased (P<0.001). Age, gender, BMI, surgical method and incidence of PGD after operation exerted no significant effect on the dynamic changes of absolute values of CD3+ CD4+ and CD3+ CD8+ T lymphocytes (all P>0.05). Primary disease before lung transplantation exerted no effect on the changes of CD3+ CD4+ T lymphocytes, whereas the postoperative absolute value of CD3+ CD8+ T lymphocytes was higher in recipients with infectious lung diseases (P<0.05). Conclusions The absolute values of CD3+, CD3+ CD4+, CD3+ CD8+ T lymphocytes in recipients with stable graft status after lung transplantation are relatively low in the early stage after lung transplantation, then gradually restore, and stabilize at 6 months after operation. Dynamic changes are not associated with age, gender, BMI, surgical method and incidence of PGD after operation of recipients. [ABSTRACT FROM AUTHOR]

Objective To investigate the expression of CD4 and CD14 immune cells in serum of female patients with Behcet’s disease ( BD), and to screen out the possible pathogenic genes.Methods BD-related data set GSE61399 was retrieved through GEO database, including 9 cases of CD4 immune cell samples and 8 cases of CD14 immune cell samples from BD patients ( experimental group), and 3 cases of CD4 immune cell samples and 9 cases of CD14 immune cell samples from healthy controls ( control group) . The limma R package was used to screen out the differentially expressed genes, KEGG database and Reactom database were used for joint pathway enrichment analysis of the differentially expressed genes, and GO database was used for GO biology process ( GOBP) and GO cellular component ( GOCC) analysis of the differentially expressed genes. STRING database was used to construct a network ( PPI) of differential genes and screened out the core genes with high node degree ( ≥10 as the standard) . Results There were 1 053 differentially expressed genes in CD14 immune cells between the experimental group and control group, including 506 up-regulated genes and 507 down-regulated genes. Two differentially expressed genes were all down-regulated in CD4 immune cells. These differentially expressed genes were more involved in membrane transport pathways. GOBP showed that the differential genes were mainly involved in biological processes such as proteasome catabolism, RNA transport, localization and nucleic acid transport. GOCC showed that the differentially expressed genes were involved in the construction of nuclear membrane, and the core genes related to BD were RPS27 A, GTPBP4, NDUFAB1, MRPL18, NIFK, MRPS5, and MRPL46, respectively.Conclusions There are differentially expressed genes in CD4 and CD14 immune cells in BD patients. These genes are mainly involved in the membrane transport pathways and vesicle-mediated transport pathways, in the biological processes such as proteasomal catabolic, RNA splicing and localization, and in the nuclear membrane composition, as well as binding, modification, and activation of key enzymes. Among the differentially expressed genes, the core genes related to the pathogenesis of BD mainly include RPS27 A, GTPBP4, NDUFAB1, MRPL18, NIFK, MRPS5, and MRPL46. [ABSTRACT FROM AUTHOR]

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Objective To identify whether regulation of Cathepsin G (CatG) expression can influence CD4+ T lymphocytes (T cells) activation in non-obese diabetic (NOD) mice, and to demonstrate the role of CatG inhibition in prevention and treatment for type 1 diabetes mellitus (T1DM). Methods A total of 36 female NOD mice (6 weeks old) were randomly divided into 3 groups according to the duration and blood glucose: normal control group, pre-diabetes (pre-DM) group, and diabetes (DM) group. CatG gene expression and CD4+ T cells activation were examined in these three groups of NOD mice (12 mice in each group). After modelling, CatG small interfering RNA (siRNA) was administrated to treat the other pre-diabetic NOD mice, and CatG inhibitor was administrated to treat diabetic NOD mice（12 mice in each intervention group）. Also control groups (12 mice in each group) were respectively set. After 8 weeks, all the mice were executed and histological analysis were performed. The effect of CatG inhibitor was investigated in NOD mice on CD4+ T cells activation, islet β cell function, and islet inflammation. Furthermore, NOD mice were injected with CatG siRNA in early stage to observe the effect of CatG knockdown on CD4+ T cells activation status and on diabetes progression. The data of three groups were analyzed with single factor analysis of variance, and the data of two groups were compared with independent samples t test. Results (1) The gene level of CatG of the 3 groups was 0.42±0.21, 0.60±0.33, 0.94±0.45, respectively (F=12.89, P< 0.05). The gene level of CatG in pre-DM group and DM group was significantly higher than that in normal control group. Meanwhile, compared with normal control group, the CD4+ T cells in spleen and peripheral blood were gradually activated in pre-DM group and DM group, resulting in more T helper (Th)1 cells and fewer Th2 and regulatory T(Treg) cells (F=11.85-21.36, all P<0.01). (2) After administration with CatG inhibitor, the level of blood glucose was decreased (t=11.26-12.45, all P<0.01) and insulin was increased (t= 11.89-13.78, all P<0.01);Meanwhile, the activation of CD4+ T cells was decreased, and the difference was statistically significant (all P<0.05）. (3) Compared with the control group, after early application of CatG siRNA, the percentage of Th1 cells was decreased [peripheral blood:(5.1±1.4)% vs (2.4±0.3)%, spleen: （10.6±2.0）% vs（6.3±1.5）%, t=20.78, 27.96, all P<0.05]. The percentage of Th2 and Treg cells was increased (t=20.59-30.25, all P<0.05). Meanwhile, the mice could maintain in normal glucose status or pre-DM status, and the insulin level could be enhanced significantly in CatG siRNA group (t=31.69-33.98, all P<0.01). Conclusion CatG is the major effector protease in CD4+ T cells activation in the pathogenesis of T1DM. Targeted inhibition of CatG may delay or block T1DM progression through alleviating CD4+ T cells activation. [ABSTRACT FROM AUTHOR]

Objective: To establish a method for isolation of T lymphocytes from peripheral blood mononuclear cells (PBMCs) by flow cytometry, and evaluate the isolation rate and survival rate. Methods: PBMCs were prepared using the human peripheral blood lymphocyte separation liquid based on gradient centrifugation. CD4+, CD8+T lymphocytes were sorting from PBMCs by flow cytometry, and the purity of these isolated cells was measured by flow cytometry and the survival rate of these cells was detected using trypan blue staining. Results: By this method, we effectively separated human peripheral blood CD4+and CD8+T lymphocytes. Without sorting by flow cytometry, the purity of CD4+lymphocytes is(50.5±11.5)%, and the purity of CD8+T lymphocytes is(15.4±7.1)%. Sorting by flow cytometry dramatically increased the purity of CD4+and CD8+T to(94.3±1.3)%, and 93.6+1.6%, respectively. The survival rate was(95.3±1.8%) for the sorted CD4+T lymphocyte, and(94.8±1.5%) for the CD8+T lymphocyte. Conclusion: This study demonstrated that flow cytometry is highly efficient in the isolation of CD4+and CD8+T lymphocytes from PBMCs, providing a guarantee for further study using these cells. Other T lymphocytes subsets also can be highly efficient isolated from PBMCs when it were labeled with different fluorescent antibodies. [ABSTRACT FROM AUTHOR]

Objective Analyze the correlation between level of CD4 cells, viral load and oral candidiasis opportunistic infections in AIDS patients, in order to deepen the understanding and provide the basis for clinical diagnosis and treatment. Methods AIDS patients of our hospital from June 2010 to June 2012 was selected. The HIV viral load was analyzed using real-time PCR test, CD3, CD4 and CD8 lymphocytes counts were detected by flow cytometry, and the oral candida strains were cultured and identified. Results Among the 85 cases with CD4+ <200 cell/L, 39 cases had oral candidiasis infection, accounting for 39%. The most common infection was （33%). and infection accounted for 0.3% and 0.1%, respectively. Plasma HIV viral load was 103~104 in 46 cases of 100 patients, with 24 cases of oral candidiasis infection, accounting for 52.2% (24/46). HIV viral load was > 105 in 22 cases, with 12 cases of oral candidiasis infection, accounting for 54.5% (12/22). Conclusion Prevalence of oral candida infections in AIDS patients has a close correlation with the CD4 cell count and viral load. [ABSTRACT FROM AUTHOR]