初步探讨甲肝疫苗诱导的细胞免疫

发布时间：2018-06-14 06:02

本文选题：甲型肝炎 + 疫苗　；参考：《北京生物制品研究所》2007年硕士论文

【摘要】： 甲型肝炎疫苗在甲型肝炎的预防和控制中发挥了积极作用。目前，甲型肝炎全病毒灭活苗在国内外已广泛使用；甲型肝炎减毒苗在国内也已得到广泛使用。实验研究和临床应用效果显示两种疫苗均具有较好的免疫原性，其免疫接种后的抗体阳转率、抗体滴度以及抗体保护的持久性等方面，已经得到充分的肯定。近年来随着对分子水平免疫反应认识的深入和基础研究的不断加强，以及方法学的进一步发展，人们评价一种疫苗的免疫应答效果，不再仅局限于体液免疫方面，而是越来越关注疫苗诱导的细胞免疫的重要作用。本研究主要评价目前国内普遍应用的两种甲肝疫苗，即甲肝减毒活疫苗与甲肝灭活疫苗，免疫接种后诱导的细胞免疫应答效果以及两者间差别，同时检测甲肝抗体滴度和抗体阳转率，探讨细胞免疫与体液免疫的关系及细胞免疫在早期抗甲肝病毒感染保护中的作用。我们从18～22岁的健康志愿者中，选择甲肝总抗体阴性者和既往甲肝疫苗接种者共18人为本次研究对象。随机分为4组，其中实验组分为2组，每组7人，分别接种甲肝减毒活疫苗和甲肝灭活疫苗；对照组中，设立阳性对照(既往有甲肝疫苗接种史，自愿加强一剂甲肝疫苗)和阴性对照(实验结束时，再接种甲肝疫苗)，每组2人。分别在0周(免疫接种前)和免疫接种后1周、2周、3周、4周、5周、6周、24周(6个月)、25周(灭活苗加强后1周)各采血一次，分离血清，以化学发光法对特异性甲肝抗体进行定性和定量检测；同时分离外周血单个核细胞PBMC，在体外将PBMC以特异性甲肝抗原刺激后，ELISPOT法检测效应T细胞分泌Th1型细胞因子IFN-γ、IL-2情况，胞内染色法测定CD3／CD4~+T淋巴细胞、CD3／CD8~+T淋巴细胞中分泌IFN-γ的阳性细胞百分比，Luminex法检测细胞培养上清中IFN-γ、IL-2、IL-10、IL-4等多种细胞因子水平。实验数据提示，应用ELISPOT、胞内染色、Luminex等检测方法，特异性好、灵敏度高，结果稳定、可靠，适用于疫苗接种后特异性细胞免疫应答的评价。实验结果显示，在免疫接种甲肝灭活疫苗、甲肝减毒活疫苗后，，早期便可诱导产生病毒特异性T细胞的细胞免疫应答反应，并且伴随高水平的效应细胞因子IFN-γ产生。同时提示，在早期的某些时间点上，两种苗所诱导的病毒特异性细胞免疫反应强度存在差别，推测与两种疫苗的甲肝抗原含量以及减毒苗活病毒在体内复制增殖后产生大量内源性抗原有关。研究数据还显示，加强接种后激发免疫记忆性在甲肝疫苗接种者是存在的。在病毒特异性T细胞增殖反应的存在条件下，接种者受到病毒性抗原的刺激下会发生快速的免疫应答反应，结果导致快速高水平的抗体和IFN-γ等抗病毒细胞因子产生。本研究初步建立了甲型肝炎疫苗的细胞免疫评价方法，通过对甲肝疫苗在人体免疫应答效果的探讨和分析，补充和完善了甲肝疫苗的免疫效果评价资料；通过人体诱导的细胞免疫的相关研究，填补了国内外关于甲肝减毒活疫苗在人体诱导的细胞免疫相关研究的空缺，为今后更加全面的评价甲肝疫苗诱导的细胞免疫以及甲肝疫苗能否用于甲肝暴露后预防提供了一定的参考依据。
[Abstract]:Hepatitis A vaccine has played a positive role in the prevention and control of hepatitis A . At present , the inactivated vaccine of hepatitis A virus has been widely used at home and abroad ;
In recent years , with the deepening of the understanding of molecular level immune response , the increasing of antibody titer and the persistence of antibody protection , it has been fully affirmed that in recent years , with the deepening of the understanding of molecular level immune response and the persistence of antibody protection , it has been evaluated that the immune response of a vaccine is no longer limited to humoral immunity , but is more and more concerned with the important role of vaccine - induced cellular immunity .

In this study , two kinds of hepatitis A vaccine , namely hepatitis A live attenuated live vaccine and hepatitis A virus inactivated vaccine , were evaluated in this study . The cellular immune response effect induced after immunization and the difference between them were evaluated . Meanwhile , the relationship between antibody titer and antibody positive rate was detected , and the relationship between cellular immunity and humoral immunity and the role of cellular immunity in the protection of early hepatitis A virus infection were discussed .

In the healthy volunteers from 18 to 22 years of age , 18 were randomly divided into 4 groups , in which the experimental group was divided into 4 groups , in which the experimental group was divided into 2 groups , 7 in each group , respectively inoculated with hepatitis A virus attenuated live vaccine and hepatitis A inactivated vaccine ;
In the control group , positive control was established ( prior to vaccination history of hepatitis A vaccine , voluntary enhancement of a dose of hepatitis A vaccine ) and negative control ( at the end of the experiment ) , 2 persons in each group . After 0 weeks ( pre - vaccination ) and 1 week after immunization respectively , 2 weeks , 3 weeks , 4 weeks , 5 weeks , 6 weeks , 24 weeks ( 6 months ) , 25 weeks ( 1 week after inactivated vaccine ) , the serum was isolated , and the specific hepatitis A antibodies were qualitatively and quantitatively detected by chemiluminescence method ;
The levels of IFN - 纬 , IL - 2 , IL - 10 and IL - 4 were measured by Luminex method .

The results showed that after immunization with hepatitis A virus inactivated vaccine and hepatitis A virus attenuated live vaccine , the cell immune response of virus - specific T cells could be induced .

In this study , the cellular immunity evaluation method of hepatitis A vaccine was established , and the evaluation data of the immune response of hepatitis A vaccine was supplemented and improved by discussing and analyzing the effect of hepatitis A vaccine on human immune response .
The study of cell immunity induced by human body fills the vacancy of the research on cell immunity induced by hepatitis A attenuated live vaccine at home and abroad , and provides a certain reference for the future comprehensive evaluation of the cell immunity induced by hepatitis A vaccine and whether the hepatitis A vaccine can be used for preventing hepatitis A exposure .
【学位授予单位】：北京生物制品研究所
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

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