Objective: To investigate the proteasome inhibitor MG132-induced apoptosis pathway in HL-60 cells and the role of allogeneic mixed lymphocyte reaction. Methods: Cell apoptosis was analyzed by flow cytometry. The ex...Objective: To investigate the proteasome inhibitor MG132-induced apoptosis pathway in HL-60 cells and the role of allogeneic mixed lymphocyte reaction. Methods: Cell apoptosis was analyzed by flow cytometry. The expressions of p21 protein, p27 protein and p53 protein in HL-60 ceils treated with MG132 were measured by Western blot. The proliferation of, peripheral blood mononuclear cells (PBMNCs) after treatment with 75 Gy irradiated HL-60 cells treated with MG132 was measured with CCK-8. Results: High-dose MG132 induced apoptosis in HL-60 cells. No significant change was observed in MG132-induced apoptosis after inhibiting caspase-8 and caspase-9 pathway. The expressions of p21 protein and p27 protein increased in MG132-induced apoptosis. HL-60 cells treated with low-dose MG132 improved the proliferation of PBMNCs from healthy volunteers. Conclusion: High-dose MG132 induced apoptosis and directly killed HL-60 ceils. MG132 induced apoptosis in a caspase-8- and caspase-9-independent pathway, p21 protein and p27 protein were involved in MG132-induced apoptosis in HL-60 cells. HL-60 cells treated with Low-dose MG132 improved the effect of promoting the proliferation of PBMNCs from healthy volunteers.展开更多
The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation i...The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation in vivo. We investigated AMLR in patients with acute- and chronic myeloid leukemia (AML and CML). AMLR was found to be significantly depressed (P<0.001) in AML patients (n=17, cpm=532±95) and CML patients (n=13, cpm=688±99) when compared with that of their healthy HLA-identical siblings serving as controls (n=17, cpm=4152±619 and n=13 cpm=4086±421, respectively). In order to understand the cellular basis of the defective AMLR in patients with AML end CML, we performed mitogen-treated T-cell cultures analysis of T-cell subsets and HLA-Ⅱ antigen detection on monocytes. The results indicated that the defect of AMLR in patients resided at the stimulator monocyte level rather than at the responder T-cell level. Enumeration of monocytes reactive with monoclonal antibody Tu22, which recognizes determinants of HLA-DQ, demonstrated that ML patients had a significantly decreased (P<0.091) number of circulating Tu22+ monocytes when compared with normal controls. These studies suggest that a deficiency of HLA-DQ+ monocytes contributes to the depression of AMLR in ML and possibly underlies the abnormalities of immune response present in this disease.展开更多
The frequency of T cells that can respond to alloantigens is unusually high.It remains unclear how T cells would respond when stimulated by multiple major histocompatibility complex(MHC)disparate alloantigens in the s...The frequency of T cells that can respond to alloantigens is unusually high.It remains unclear how T cells would respond when stimulated by multiple major histocompatibility complex(MHC)disparate alloantigens in the same cultures.In this report,we examined potential interactions of T cell clones that were stimulated simultaneously by two sets of complete MHC disparate alloantigens using mixed lymphocyte reaction(MLR).In this assay,we observed that proliferation of B6 lymphocytes(H-2b)stimulated by both BALB/c(H-2d)and C_(3)H(H-2k)allogeneic cells was not increased but rather reduced as compared to B6 cells stimulated with either BALB/c or C_(3)H allogeneic cells.Interestingly,interleukin(IL)-10 expressions at both protein level and mRNA level was signifi cantly increased in cultures stimulated with the two MHC alloantigens,while IL-2,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β1 production did not show any differences.In addition,Foxp_(3) mRNA expression was comparable amongst all groups.In conclusion,we observed an inhibitory effect in T cell proliferation in response to multiple MHC mismatched alloantigens in MLR,and this effect might be associated with the upregulation of IL-10 expression.展开更多
目的:研究恩替卡韦(ETV)对慢性乙型肝炎(CHB)患者外周血树突状细胞(dendritic cell,DC)功能的影响.方法:体外常规分离CHB患者及健康人外周血单个核细胞,诱导扩增后常规培养.第4天将其与一定浓度的恩替卡韦共培养,第8天收获DC进行细胞表...目的:研究恩替卡韦(ETV)对慢性乙型肝炎(CHB)患者外周血树突状细胞(dendritic cell,DC)功能的影响.方法:体外常规分离CHB患者及健康人外周血单个核细胞,诱导扩增后常规培养.第4天将其与一定浓度的恩替卡韦共培养,第8天收获DC进行细胞表型、同种异体混合淋巴细胞反应等相关检测.结果:细胞培养8 d时DC形态分化健康对照组优于CHB ETV处理组,CHB ETV处理组优于CHB组;CHB组CD1a(35.73±3.12 vs 62.31±5.22,P<0.01),CD80(28.19±1.64 vs 45.38±3.10,P<0.01),CD83(22.24±2.14 vs 40.63±7.21,P<0.01)及HLA-DR(36.74±0.98 vs 56.05±3.89,P<0.01)表达明显低于健康对照组,而ETV处理组与CHB组相比CD83 (27.41±9.23 vs 22.24±2.14,P<0.05),CD80(32.67±7.82 vs 28.19±1.64,P<0.05)及HLA-DR(40.84±5.57 vs 36.74±0.98,P<0.01)显著高表达;淋巴细胞增殖能力测定ETV处理组DC刺激同种异体T淋巴细胞增殖能力较CHB组增强(1.53±0.09 vs 1.45±0.12,P<0.05).结论:恩替卡韦作为治疗CHB的新一代核苷类药物,除了直接抑制乙肝病毒DNA合成外,也能够增强CHB患者外周血DC的功能,通过调节机体的免疫系统发挥间接抗病毒作用.展开更多
基金supported by the grants from Science and Research Special Foundation of Wuhan University of Science and Technology(No.zx0802)the Natural Science Foundation of Jiangxi Province(0640190)
文摘Objective: To investigate the proteasome inhibitor MG132-induced apoptosis pathway in HL-60 cells and the role of allogeneic mixed lymphocyte reaction. Methods: Cell apoptosis was analyzed by flow cytometry. The expressions of p21 protein, p27 protein and p53 protein in HL-60 ceils treated with MG132 were measured by Western blot. The proliferation of, peripheral blood mononuclear cells (PBMNCs) after treatment with 75 Gy irradiated HL-60 cells treated with MG132 was measured with CCK-8. Results: High-dose MG132 induced apoptosis in HL-60 cells. No significant change was observed in MG132-induced apoptosis after inhibiting caspase-8 and caspase-9 pathway. The expressions of p21 protein and p27 protein increased in MG132-induced apoptosis. HL-60 cells treated with low-dose MG132 improved the proliferation of PBMNCs from healthy volunteers. Conclusion: High-dose MG132 induced apoptosis and directly killed HL-60 ceils. MG132 induced apoptosis in a caspase-8- and caspase-9-independent pathway, p21 protein and p27 protein were involved in MG132-induced apoptosis in HL-60 cells. HL-60 cells treated with Low-dose MG132 improved the effect of promoting the proliferation of PBMNCs from healthy volunteers.
文摘The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation in vivo. We investigated AMLR in patients with acute- and chronic myeloid leukemia (AML and CML). AMLR was found to be significantly depressed (P<0.001) in AML patients (n=17, cpm=532±95) and CML patients (n=13, cpm=688±99) when compared with that of their healthy HLA-identical siblings serving as controls (n=17, cpm=4152±619 and n=13 cpm=4086±421, respectively). In order to understand the cellular basis of the defective AMLR in patients with AML end CML, we performed mitogen-treated T-cell cultures analysis of T-cell subsets and HLA-Ⅱ antigen detection on monocytes. The results indicated that the defect of AMLR in patients resided at the stimulator monocyte level rather than at the responder T-cell level. Enumeration of monocytes reactive with monoclonal antibody Tu22, which recognizes determinants of HLA-DQ, demonstrated that ML patients had a significantly decreased (P<0.091) number of circulating Tu22+ monocytes when compared with normal controls. These studies suggest that a deficiency of HLA-DQ+ monocytes contributes to the depression of AMLR in ML and possibly underlies the abnormalities of immune response present in this disease.
基金supported by a grant from the National Nature Science Foundation of China(NSFC81302548)to J Zhou.
文摘The frequency of T cells that can respond to alloantigens is unusually high.It remains unclear how T cells would respond when stimulated by multiple major histocompatibility complex(MHC)disparate alloantigens in the same cultures.In this report,we examined potential interactions of T cell clones that were stimulated simultaneously by two sets of complete MHC disparate alloantigens using mixed lymphocyte reaction(MLR).In this assay,we observed that proliferation of B6 lymphocytes(H-2b)stimulated by both BALB/c(H-2d)and C_(3)H(H-2k)allogeneic cells was not increased but rather reduced as compared to B6 cells stimulated with either BALB/c or C_(3)H allogeneic cells.Interestingly,interleukin(IL)-10 expressions at both protein level and mRNA level was signifi cantly increased in cultures stimulated with the two MHC alloantigens,while IL-2,tumor necrosis factor(TNF)-α,transforming growth factor(TGF)-β1 production did not show any differences.In addition,Foxp_(3) mRNA expression was comparable amongst all groups.In conclusion,we observed an inhibitory effect in T cell proliferation in response to multiple MHC mismatched alloantigens in MLR,and this effect might be associated with the upregulation of IL-10 expression.
文摘目的:研究恩替卡韦(ETV)对慢性乙型肝炎(CHB)患者外周血树突状细胞(dendritic cell,DC)功能的影响.方法:体外常规分离CHB患者及健康人外周血单个核细胞,诱导扩增后常规培养.第4天将其与一定浓度的恩替卡韦共培养,第8天收获DC进行细胞表型、同种异体混合淋巴细胞反应等相关检测.结果:细胞培养8 d时DC形态分化健康对照组优于CHB ETV处理组,CHB ETV处理组优于CHB组;CHB组CD1a(35.73±3.12 vs 62.31±5.22,P<0.01),CD80(28.19±1.64 vs 45.38±3.10,P<0.01),CD83(22.24±2.14 vs 40.63±7.21,P<0.01)及HLA-DR(36.74±0.98 vs 56.05±3.89,P<0.01)表达明显低于健康对照组,而ETV处理组与CHB组相比CD83 (27.41±9.23 vs 22.24±2.14,P<0.05),CD80(32.67±7.82 vs 28.19±1.64,P<0.05)及HLA-DR(40.84±5.57 vs 36.74±0.98,P<0.01)显著高表达;淋巴细胞增殖能力测定ETV处理组DC刺激同种异体T淋巴细胞增殖能力较CHB组增强(1.53±0.09 vs 1.45±0.12,P<0.05).结论:恩替卡韦作为治疗CHB的新一代核苷类药物,除了直接抑制乙肝病毒DNA合成外,也能够增强CHB患者外周血DC的功能,通过调节机体的免疫系统发挥间接抗病毒作用.