rHBsAg刺激下乙肝疫苗无应答者离体PBMC的CD28、PD-1等表达量观察  

Expression levels of CD28 and PD-1 in isolated PBMC of non-responders to hepatitis B vaccine stimulated by rHBsAg

作　　者：邵美琪 刘晓光 苏百乔 万莹 高金霞[1] 王栋[1] 牛莹莹[1] 范兴君[1] 靳革[1] 唐颖 谷红梅[1] SHAO Mei-qi(Department of Public Health,Mudanjiang Medical University,Mudanjiang 157011,Heilongjiang,China)

机构地区：[1]牡丹江医学院公共卫生学院,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2024年第3期16-20,25,共6页Journal of Mudanjiang Medical University

基　　金：黑龙江省卫生健康委科研项目(2020-414);牡丹江市应用技术研究与开发计划项目(HT2022NS106)。

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。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 exp

关 键 词：乙肝疫苗 rHBsAg 共刺激分子 

分 类 号：R512.62[医药卫生—内科学]