甲乙肝联合疫苗中甲肝乙肝抗原相互作用机制的探索研究

发布时间：2018-01-15 12:35

本文关键词：甲乙肝联合疫苗中甲肝乙肝抗原相互作用机制的探索研究　出处：《中国协和医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 甲型肝炎(甲肝)和乙型肝炎(乙肝)是最常见的病毒性肝炎,它们是严重危害公众健康的传染病。其中乙肝病毒(hepatitis B viral)感染可以发展成慢性肝病、肝硬化或肝细胞癌。甲肝病毒(hepatitis A viral)感染一般不会引起慢性肝病,少数成人患者病情迁延或复发,个别患者病程可达六个月以上,预后较差。尽管甲肝和乙肝的主要传播途径不同,但近年来有大量报道,肝炎患者有相当一部分人同时感染甲肝和乙肝。因此,预防甲肝和乙肝对控制病毒性肝炎的感染和流行尤为重要。正因为甲肝和乙肝对人类健康危害很大,而且目前尚无特效药物治疗,所以甲肝、乙肝疫苗的广泛应用可对甲肝乙肝病毒的流行起到非常重要的遏制作用。近年来,研制和使用含有两种或多种抗原,可以预防两种或多种疾病的联合疫苗受到世界各国的重视,已成为疫苗开发的热点。联合疫苗为儿童免疫规划的连续、简化和扩大所必需。甲乙型肝炎联合疫苗(Combined Hepatities A and Bvaccine)是具有甲、乙型肝炎双重特异性保护性能,同时预防这两种疾病的一种联合疫苗。目前,在甲乙肝联合疫苗的研究中表明,甲乙型肝炎联合疫苗诱导机体产生的抗体水平高于单价疫苗。研究者们普遍认为采用联合疫苗接种比单价苗接种能诱导机体产生更好的乙肝反应,推测两种抗原可能相互作用互为佐剂,但尚无系统的免疫学研究的报道。本实验通过对不同配比的实验性甲乙肝联合疫苗和单价苗免疫小鼠后的四个指标的检测:流式细胞仪检测细胞因子水平和T细胞亚群分型,MTT法测脾细胞增殖水平,ELISA法检测抗体水平,来评估联合疫苗的细胞及体液免疫情况,以此衡量免疫应答的性质,评估免疫记忆效果,最终获得两种疫苗的最佳配比,并以此进一步分析甲乙联合疫苗中两种抗原的相互作用机制,同时也期望更全面的了解甲乙肝联合疫苗的免疫原性。我们采用医学生物学研究所生产的甲肝灭活疫苗和深圳康泰生物制品公司生产的基因工程重组乙肝酵母疫苗半成品,以640Eu/ml甲肝抗原与10ug和20ug的乙肝表面抗原进行配比混合。不同配比的联合疫苗与单价疫苗分别于BALB/C小鼠腓肠肌接种各200μl/只。每组5只小鼠,于接种后第4周,再经腓肠肌注射相同的疫苗,加强免疫1次,200μl/只。初次免疫后1、2、3、4周和加强免疫后2周,分别检测体液和细胞免疫情况。结果表明:甲乙肝联合疫苗均显示出能刺激小鼠产生较单价苗更强的免疫应答。在初次免疫的早期,加入甲肝抗原在一定程度上增强了乙肝疫苗的细胞免疫原性,起到了佐剂的作用。综合分析本试验的结果,我们得出的结论是我所自主研制的甲乙肝联合疫苗640Eu/ml:10μg/ml配比就免疫效果来说是适合需要的。联合疫苗能刺激小鼠产生较强的Th1应答,加强免疫后,Th1及Th2型细胞因子均显著增加。
[Abstract]:Hepatitis A virus (HAV) and hepatitis B virus (HBV) is the most common type of viral hepatitis, which is a serious public health hazard of infectious diseases. The hepatitis B virus (hepatitis B viral) infection can develop into chronic liver disease, cirrhosis or hepatocellular carcinoma. Hepatitis A virus (hepatitis A viral) infection generally does not cause chronic liver disease a few adult patients with persistent disease or relapse, individual patients up to six months of poor prognosis. Although different main transmission way of hepatitis A and B, but in recent years a large number of reported hepatitis patients with quite a few people infected with hepatitis A and B at the same time. Therefore, the prevention of hepatitis A and B is particularly important for control of viral hepatitis the infection and prevalence of hepatitis A and B. Because of great harm to human health, but there is no effective drug treatment, so the hepatitis A, hepatitis B vaccine can be widely applied to hepatitis A hepatitis B disease The popular poison to restrain is very important. In recent years, the development and use of containing two or more antigens, vaccine preventable diseases of two or more attention all over the world, has become a hot topic in vaccine development. Combined vaccine for continuous childhood immunization, to simplify and expand the necessary joint armor. Hepatitis B vaccine (Combined Hepatities A and Bvaccine) is a dual specificity, hepatitis B protection performance, a combined vaccine of these two diseases. At present, the research shows that in a combined with hepatitis B vaccine, the antibody level of combined hepatitis A and B vaccine induce higher than monovalent vaccine researchers. Is generally believed that hepatitis B vaccine was used than monovalent vaccine inoculation can induce a better, that two antigens may interact each other as adjuvant, but there is no free system A report on the Phytophthora study.
In this experiment, through the detection of four indexes of different ratio of experimental hepatitis A and B combined vaccine and monovalent vaccine immunized mice after the detection of cytokines levels and T cell subset by flow cytometry, measuring the spleen cell proliferation level by MTT, detection of the antibody level of ELISA method to evaluate the cellular and humoral immunity of contact vaccine, nature as a measure of immune response, immune memory effect assessment, finally get the best ratio of the two kinds of vaccines, and further analysis of the interaction mechanism of two antigens in B combined vaccine, but also expect a more comprehensive understanding of a combined with hepatitis B vaccine immunogenicity.
We use the Institute of biological medicine production of inactivated hepatitis A vaccine and Shenzhen Kangtai biological products company production of recombinant yeast hepatitis B vaccine semi-finished products, mix with the hepatitis B surface antigen 640Eu/ml antigen with 10ug and 20ug. The different ratio of combined vaccine and monovalent vaccine in BALB/C mice inoculated with gastrocnemius muscle 200 l/. 5 mice in each group, at fourth weeks after inoculation, the gastrocnemius muscle injected with the same vaccine immunization for 1 times, 200 l/. 1,2,3,4 weeks after the first immunization and strengthening immunity after 2 weeks, were used to detect the humoral and cellular immune condition. The results show that a combined with hepatitis B vaccine showed that stimulation of mouse immune response than monovalent vaccine. More early in the first immunization, with enhanced cellular immune antigen of hepatitis B vaccine immunogenicity in a certain extent, played the role of adjuvant comprehensive. Analysis of the results, we concluded that I developed a 640Eu/ml:10 g/ml combined with hepatitis B vaccine immune effect ratio is suitable for the needs of the joint. The vaccine can stimulate mice to produce a strong Th1 response, strengthen the immunity, Th1 and Th2 type cytokines were significantly increased.

【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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