【摘要】： 在世界范圍內(nèi)沙眼衣原體(Chlamydia trachomatis,C.t)是細(xì)菌性性傳播疾病的首要原因,盡管現(xiàn)在可以得到各種抗生素,但是每年大約還有10億人被臨床診斷為沙眼衣原體感染。大多數(shù)衣原體生殖道感染起初是沒有癥狀、沒有治療從而可能導(dǎo)致嚴(yán)重的并發(fā)癥,如盆腔炎、異位妊娠和不孕,給社會經(jīng)濟(jì)帶來了嚴(yán)重的負(fù)擔(dān)。因此,我們需要在世界范圍內(nèi)控制沙眼衣原體感染,而最好的方法是有效的疫苗來預(yù)防沙眼衣原體導(dǎo)致的性傳播疾病。盡管幾個世紀(jì)的努力,但還是沒有成功的疫苗問世。E型沙眼衣原體主要引起性傳播感染,最近研究者們主要研究衣原體主要外膜蛋白作為疫苗的候選抗原。E型沙眼衣原體重組主要外膜蛋白疫苗(rMOMP)和DNA疫苗是世界范圍內(nèi)的研究熱點(diǎn),因此,我們實(shí)驗(yàn)室研制出這兩種疫苗并研究他們在小鼠體內(nèi)誘發(fā)的免疫效應(yīng)。 目的：表達(dá)感染泌尿生殖道的C.t優(yōu)勢型—E型主要外膜蛋白rMOMP,連同本實(shí)驗(yàn)室構(gòu)建保存的MOMP蛋白的DNA,研究并比較每種疫苗各自單獨(dú)免疫和DNA疫苗蛋白疫苗聯(lián)合免疫在小鼠體內(nèi)誘發(fā)的免疫應(yīng)答和免疫保護(hù)作用。 方法：以BALB/c小鼠為實(shí)驗(yàn)動物,采用肌肉注射和陰道涂布的免疫途徑,將小鼠分為rMOMP佐劑肌注組、rMOMP肌注組、DNA肌注組、rMOMP陰道組、DNA陰道組、DNA聯(lián)合rMOMP組、PBS肌注陰性對照組、PBS陰道陰性對照組和滅活C.t E型原體(EB)肌注陽性對照組。通過血清和生殖道局部抗體檢測、細(xì)胞因子檢測、抗體中和試驗(yàn)、遲發(fā)型超敏反應(yīng)、脾淋巴細(xì)胞增殖試驗(yàn)、實(shí)驗(yàn)動物經(jīng)陰道途徑進(jìn)行C.t E型攻擊后的C.t清除情況的觀察來研究C.t E型rMOMP疫苗和DNA疫苗的免疫效果。 結(jié)果：(1)C.t E型rMOMP蛋白疫苗和DNA疫苗通過肌注途徑單獨(dú)免疫的小鼠能夠誘導(dǎo)產(chǎn)生一定的C.t特異性細(xì)胞免疫和體液免疫,對C.t E型生殖道的攻擊具有保護(hù)作用,而且rMOMP蛋白疫苗的免疫效果優(yōu)于DNA疫苗。(2) rMOMP蛋白疫苗和弗氏佐劑聯(lián)合免疫能夠顯著增強(qiáng)CtE型rMOMP蛋白疫苗所誘導(dǎo)的免疫反應(yīng)。(3)DNA疫苗和rMOMP蛋白疫苗聯(lián)合免疫組誘導(dǎo)的免疫效應(yīng)顯著優(yōu)于rMOMP蛋白疫苗和DNA疫苗單獨(dú)免疫組。(4)通過陰道免疫途徑,無論是蛋白疫苗還是DNA疫苗都沒有誘導(dǎo)小鼠產(chǎn)生特異性免疫應(yīng)答。 結(jié)論：C.t E型rMOMP蛋白疫苗和DNA疫苗通過肌注免疫能夠在實(shí)驗(yàn)小鼠體內(nèi)發(fā)揮一定的免疫保護(hù)作用,且rMOMP蛋白疫苗的免疫效果優(yōu)于DNA疫苗,rMOMP和DNA聯(lián)合免疫能夠顯著增強(qiáng)免疫效應(yīng),通過陰道涂布的方式不能產(chǎn)生保護(hù)性免疫應(yīng)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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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