【摘要】 目的 观察乙肝疫苗无应答者离体外周血单个核细胞(peripheral blood mononuclear cells, PBMC)在重组乙肝表面抗原(recombinant hepatitis B surface antigen, rHBsAg)刺激下分化抗原28(cluster of differentiation 28,CD28)及程序性死亡受体-1(programmed death-1,PD-1)等共刺激分子表达量与乙肝疫苗有应答者的差异，为探索提升乙肝疫苗无应答者的应答能力提供一定数据参考。方法 通过前期问卷调查、志愿者乙肝病毒血清学标准物(HBV serum marker, HBV-M)检测结果筛选，随机抽取30名符合条件的乙肝疫苗无应答者入组，并随机抽取30名乙肝疫苗有应答者作为对照进入实验。对乙肝疫苗无应答者和有应答者的离体PBMC同步进行rHBsAg体外刺激，并使用流式细胞仪对T细胞亚群及其表面共刺激分子CD28、细胞毒性T淋巴细胞相关蛋白4(cytotoxic T lymphocyte associated protein 4,CTLA4)、T细胞免疫球蛋白黏蛋白3(T cell immunoglobulin mucin 3,Tim3)及PD-1表达量检测。所有测量数据校对后使用SPSS 23.0对乙肝疫苗无应答组与有应答组行T细胞亚群及共刺激分子表达量比较。结果 乙肝疫苗无应答组与有应答组平均年龄、性别构成差异无统计学意义；相较于乙肝疫苗有应答者，乙肝疫苗无应答者离体PBMC初始CD8~+PD-1~+表达量略高(z=2.775,P=0.006),在rHBsAg刺激下，T细胞亚群CD3~+、CD4~+、CD8~+、CD4~+/CD8~+的表达量差异无统计学意义，共刺激分子CD4~+CD28~+、CD4~+CTLA4~+、CD8~+CTLA4~+、CD4~+PD-1~+、CD4~+Tim3~+、CD8~+Tim3~+的表达量差异无统计学意义，CD8~+CD28~+的表达量低(t=2.606,P=0.012),CD8~+PD-1~+的表达量偏高(z=2.339,P=0.019)。结论 乙肝疫苗有应答及无应答两类人群外周血T细胞PD-1~+表达量存在一定差异，能否通过调节PD-1~+表达量提升乙肝疫苗无应答者的应答能力有待进一步研究。更多还原

【Abstract】 Objective To observe the differences in the expression levels of cluster of differentiation 28(CD28) and programmed death-1(PD-1)and other co-stimulatory molecules in vitro peripheral blood mononuclear cells(PBMC) stimulated by recombinant hepatitis B surface antigen(rHBsAg) between non-responders to hepatitis B vaccine and those who responded to hepatitis B vaccine, which provides some data reference for exploring and improving the response-ability of non-responders to hepatitis B vaccine.Methods Based on the preliminary questionnaire survey and the detection results of HBV serum marker(HBV-M),thirty eligible hepatitis B vaccine non-responders and thirty hepatitis B vaccine responders were randomly selected as controls to enter the experiment.In vitro, rHBsAg stimulation was performed on PBMC of both non-responders and responders to hepatitis B vaccine, and the expression levels of T cell subpopulations and their surface co-stimulatory molecules CD28,cytotoxic T lymphocyte-associated protein 4(CTLA4),T cell immunoglobulin mucin 3(Tim3) and PD-1were detected by flow cytometry.After collating all the measured data, the expression of T cell subsets and co-stimulatory molecules were compared between the non-responsive group and the responsive group using SPSS23.0.Results There was no significant difference in mean age and gender composition between the non-responding group and the responding group.Compared with those who responded to hepatitis B vaccine, the initial expression of CD8~+PD-1~+ in isolated PBMC was slightly higher in those who did not respond to hepatitis B vaccine(z=2.775,P=0.006).Under the stimulation of rHBsAg, the expression levels of T cell subsets CD3~+,CD4~+,CD8~+,and CD4~+/CD8~+ in non-responders to hepatitis B vaccine were not statistically significant compared with those in responders.There was no significant difference in the expression levels of co-stimulatory molecules CD4~+CD28~+,CD4~+CTLA4~+,CD8~+CTLA4~+,CD4~+PD-1~+,CD4~+Tim3~+,and CD8~+Tim3~+ between non-responders and responders under rHBsAg stimulation, while the expression level of CD8~+CD28~+ was low(t=2.606,P=0.012),and CD8~+PD-1~+ was highly expressed(z=2.339,P=0.019).Conclusion There are some differences in PD-1~+ expression of peripheral T cells in the two groups of responder and non-responder to hepatitis B vaccine.Therefore, whether the response-ability of non-responders can be improved by regulating the expression of PD-1~+ expression needs further study.更多还原