E型沙眼衣原体重组主要外膜蛋白多肽疫苗免疫效应的研究与DNA疫苗的比照

发布时间：2018-08-06 10:45

【摘要】： 在世界范围内沙眼衣原体(Chlamydia trachomatis,C.t)是细菌性性传播疾病的首要原因,尽管现在可以得到各种抗生素,但是每年大约还有10亿人被临床诊断为沙眼衣原体感染。大多数衣原体生殖道感染起初是没有症状、没有治疗从而可能导致严重的并发症,如盆腔炎、异位妊娠和不孕,给社会经济带来了严重的负担。因此,我们需要在世界范围内控制沙眼衣原体感染,而最好的方法是有效的疫苗来预防沙眼衣原体导致的性传播疾病。尽管几个世纪的努力,但还是没有成功的疫苗问世。E型沙眼衣原体主要引起性传播感染,最近研究者们主要研究衣原体主要外膜蛋白作为疫苗的候选抗原。E型沙眼衣原体重组主要外膜蛋白疫苗(rMOMP)和DNA疫苗是世界范围内的研究热点,因此,我们实验室研制出这两种疫苗并研究他们在小鼠体内诱发的免疫效应。目的：表达感染泌尿生殖道的C.t优势型—E型主要外膜蛋白rMOMP,连同本实验室构建保存的MOMP蛋白的DNA,研究并比较每种疫苗各自单独免疫和DNA疫苗蛋白疫苗联合免疫在小鼠体内诱发的免疫应答和免疫保护作用。方法：以BALB/c小鼠为实验动物,采用肌肉注射和阴道涂布的免疫途径,将小鼠分为rMOMP佐剂肌注组、rMOMP肌注组、DNA肌注组、rMOMP阴道组、DNA阴道组、DNA联合rMOMP组、PBS肌注阴性对照组、PBS阴道阴性对照组和灭活C.t E型原体(EB)肌注阳性对照组。通过血清和生殖道局部抗体检测、细胞因子检测、抗体中和试验、迟发型超敏反应、脾淋巴细胞增殖试验、实验动物经阴道途径进行C.t E型攻击后的C.t清除情况的观察来研究C.t E型rMOMP疫苗和DNA疫苗的免疫效果。结果：(1)C.t E型rMOMP蛋白疫苗和DNA疫苗通过肌注途径单独免疫的小鼠能够诱导产生一定的C.t特异性细胞免疫和体液免疫,对C.t E型生殖道的攻击具有保护作用,而且rMOMP蛋白疫苗的免疫效果优于DNA疫苗。(2) rMOMP蛋白疫苗和弗氏佐剂联合免疫能够显著增强CtE型rMOMP蛋白疫苗所诱导的免疫反应。(3)DNA疫苗和rMOMP蛋白疫苗联合免疫组诱导的免疫效应显著优于rMOMP蛋白疫苗和DNA疫苗单独免疫组。(4)通过阴道免疫途径,无论是蛋白疫苗还是DNA疫苗都没有诱导小鼠产生特异性免疫应答。结论：C.t E型rMOMP蛋白疫苗和DNA疫苗通过肌注免疫能够在实验小鼠体内发挥一定的免疫保护作用,且rMOMP蛋白疫苗的免疫效果优于DNA疫苗,rMOMP和DNA联合免疫能够显著增强免疫效应,通过阴道涂布的方式不能产生保护性免疫应答.
[Abstract]:Chlamydia trachomatis (Chlamydia trachomatisC. T) is the leading cause of bacterial sexually transmitted diseases worldwide. Although antibiotics are available, about 1 billion people are clinically diagnosed with chlamydia trachomatis infection every year. Most chlamydia genital tract infections are initially asymptomatic and without treatment can lead to serious complications such as pelvic inflammatory disease ectopic pregnancy and infertility which impose a severe socio-economic burden. Therefore, we need to control chlamydia trachomatis infection worldwide, and the best way is to prevent STDs caused by Chlamydia trachomatis. Despite centuries of effort, there has been no successful vaccine for Chlamydia trachomatis, a major cause of sexually transmitted infections. Recently, researchers have mainly studied the main outer membrane protein of chlamydia trachomatis as candidate antigen. E recombinant main outer membrane protein vaccine of Chlamydia trachomatis (rMOMP) and DNA vaccine is a worldwide research hotspot. Our lab developed these two vaccines and studied their immune effects in mice. Objective: to express the main outer membrane protein rMOMP of C.t dominant type E in genitourinary tract infection, and to study and compare each vaccine alone and the combined immunization of DNA vaccine protein vaccine. Immune response and protective effect induced in mice. Methods: BALB/c mice were used as experimental animals by intramuscular injection and vaginal coating. The mice were divided into two groups: rMOMP adjuvant intramuscular injection group (rMOMP adjuvant injection group), rMOMP intramuscular injection group (rMOMP vaginal injection group) and rMOMP group (rMOMP negative control group) and inactivated (EB) positive control group. Local antibody detection, cytokine detection, antibody neutralization test, delayed type hypersensitivity test, splenic lymphocyte proliferation test were used. To study the immune effect of C.t-E rMOMP vaccine and DNA vaccine, we observed the clearance of C. t E type rMOMP vaccine and DNA vaccine through vagina. Results: (1) mice immunized with C.t-E rMOMP protein vaccine and DNA vaccine alone by intramuscular injection could induce specific cellular and humoral immunity, which had protective effect on the attack of C.t-E type reproductive tract. Moreover, the immune effect of rMOMP protein vaccine was better than that of DNA vaccine. (2) combined immunization with rMOMP protein vaccine and Freund's adjuvant could significantly enhance the immune response induced by CtE type rMOMP protein vaccine. (3) the immune response induced by DNA vaccine and rMOMP protein vaccine combined immunization group was significantly enhanced. The immune effect was significantly better than that of rMOMP protein vaccine and DNA vaccine alone. (4) through vaginal immunization, Neither protein vaccine nor DNA vaccine could induce specific immune response in mice. Conclusion rMOMP protein vaccine and DNA vaccine can play a protective role in mice by intramuscular injection, and the immune effect of rMOMP protein vaccine is better than that of DNA vaccine rMOMP and DNA. Vaginal coating does not produce a protective immune response.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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