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  本文關(guān)鍵詞：甲乙肝聯(lián)合疫苗中甲肝乙肝抗原相互作用機制的探索研究　出處：《中國協(xié)和醫(yī)科大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 甲乙聯(lián)合疫苗 細胞免疫應(yīng)答 體液免疫應(yīng)答 相互作用

【摘要】： 甲型肝炎(甲肝)和乙型肝炎(乙肝)是最常見的病毒性肝炎,它們是嚴(yán)重危害公眾健康的傳染病。其中乙肝病毒(hepatitis B viral)感染可以發(fā)展成慢性肝病、肝硬化或肝細胞癌。甲肝病毒(hepatitis A viral)感染一般不會引起慢性肝病,少數(shù)成人患者病情遷延或復(fù)發(fā),個別患者病程可達六個月以上,預(yù)后較差。盡管甲肝和乙肝的主要傳播途徑不同,但近年來有大量報道,肝炎患者有相當(dāng)一部分人同時感染甲肝和乙肝。因此,預(yù)防甲肝和乙肝對控制病毒性肝炎的感染和流行尤為重要。正因為甲肝和乙肝對人類健康危害很大,而且目前尚無特效藥物治療,所以甲肝、乙肝疫苗的廣泛應(yīng)用可對甲肝乙肝病毒的流行起到非常重要的遏制作用。近年來,研制和使用含有兩種或多種抗原,可以預(yù)防兩種或多種疾病的聯(lián)合疫苗受到世界各國的重視,已成為疫苗開發(fā)的熱點。聯(lián)合疫苗為兒童免疫規(guī)劃的連續(xù)、簡化和擴大所必需。甲乙型肝炎聯(lián)合疫苗(Combined Hepatities A and Bvaccine)是具有甲、乙型肝炎雙重特異性保護性能,同時預(yù)防這兩種疾病的一種聯(lián)合疫苗。目前,在甲乙肝聯(lián)合疫苗的研究中表明,甲乙型肝炎聯(lián)合疫苗誘導(dǎo)機體產(chǎn)生的抗體水平高于單價疫苗。研究者們普遍認(rèn)為采用聯(lián)合疫苗接種比單價苗接種能誘導(dǎo)機體產(chǎn)生更好的乙肝反應(yīng),推測兩種抗原可能相互作用互為佐劑,但尚無系統(tǒng)的免疫學(xué)研究的報道。 本實驗通過對不同配比的實驗性甲乙肝聯(lián)合疫苗和單價苗免疫小鼠后的四個指標(biāo)的檢測:流式細胞儀檢測細胞因子水平和T細胞亞群分型,MTT法測脾細胞增殖水平,ELISA法檢測抗體水平,來評估聯(lián)合疫苗的細胞及體液免疫情況,以此衡量免疫應(yīng)答的性質(zhì),評估免疫記憶效果,最終獲得兩種疫苗的最佳配比,并以此進一步分析甲乙聯(lián)合疫苗中兩種抗原的相互作用機制,同時也期望更全面的了解甲乙肝聯(lián)合疫苗的免疫原性。 我們采用醫(yī)學(xué)生物學(xué)研究所生產(chǎn)的甲肝滅活疫苗和深圳康泰生物制品公司生產(chǎn)的基因工程重組乙肝酵母疫苗半成品,以640Eu/ml甲肝抗原與10ug和20ug的乙肝表面抗原進行配比混合。不同配比的聯(lián)合疫苗與單價疫苗分別于BALB/C小鼠腓腸肌接種各200μl/只。每組5只小鼠,于接種后第4周,再經(jīng)腓腸肌注射相同的疫苗,加強免疫1次,200μl/只。初次免疫后1、2、3、4周和加強免疫后2周,分別檢測體液和細胞免疫情況。結(jié)果表明:甲乙肝聯(lián)合疫苗均顯示出能刺激小鼠產(chǎn)生較單價苗更強的免疫應(yīng)答。在初次免疫的早期,加入甲肝抗原在一定程度上增強了乙肝疫苗的細胞免疫原性,起到了佐劑的作用。綜合分析本試驗的結(jié)果,我們得出的結(jié)論是我所自主研制的甲乙肝聯(lián)合疫苗640Eu/ml:10μg/ml配比就免疫效果來說是適合需要的。聯(lián)合疫苗能刺激小鼠產(chǎn)生較強的Th1應(yīng)答,加強免疫后,Th1及Th2型細胞因子均顯著增加。
[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.

【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R392

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本文編號：1428425