中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (7): 492-495.

• 论著 • 上一篇    下一篇

E型沙眼衣原体重组主外膜蛋白疫苗对恒河猴的免疫效应

李宜儒1,姚卫锋2,盛彩虹1,冯斌2,展小飞1,李玲杰3,尤聪2,李艳飞2,刘原君4,齐蔓莉2,刘全忠2   

  1. 1. 天津医科大学总医院
    2. 天津医科大学总医院皮肤性病科
    3. 天津医科大学总医院皮肤科
    4. 天津医科大学总医院/天津性传播疾病研究所
  • 收稿日期:2011-08-09 修回日期:2012-01-28 出版日期:2012-07-15 发布日期:2012-07-02
  • 通讯作者: 刘全忠 E-mail:liuquanzhong@medmail.com.cn
  • 基金资助:

    国家自然科学基金项目

Immune responses induced by the recombinant major outer membrane protein vaccine against Chlamydia trachomatis E serotype in rhesus monkeys

  • Received:2011-08-09 Revised:2012-01-28 Online:2012-07-15 Published:2012-07-02
  • Contact: quanzhong liu E-mail:liuquanzhong@medmail.com.cn

PDF

28

赞

摘要：

目的 探讨E型重组主外膜蛋白（rMOMP）对恒河猴诱导产生的特异性免疫应答效应。 方法 恒河猴分3组，每组2只，分别为佐剂蛋白组、佐剂组、对照组。各组均于第0（免疫起始周）、2、4周双侧肱三头肌注射蛋白疫苗、佐剂和磷酸盐缓冲液（PBS）。末次免疫后2周，酶联免疫吸附试验（ELISA）检测恒河猴血清中沙眼衣原体特异性IgG抗体和干扰素γ（IFN-γ），以及阴道冲洗液中沙眼衣原体特异性sIgA抗体，MTT法检测恒河猴淋巴细胞特异性增殖反应，观察恒河猴的迟发型超敏反应，以及恒河猴血清抗体中和试验，同源攻击后免疫荧光检测衣原体。用SPSS 14.0软件进行单向方差分析和LSD法进行组间多重比较。结果 佐剂蛋白组恒河猴血清衣原体特异性IgG抗体A405值为1.718 ± 0.213，淋巴细胞增殖指数为7.012 ± 1.026，IFN-γ水平为（1086.000 ± 121.730） ng/L，迟发型超敏反应皮丘直径为（11.000 ± 2.134） mm；佐剂组分别为0.841 ± 0.315、4.473 ± 1.850、（409.000 ± 53.440） ng/L、（3.000 ± 0.914） mm；对照组分别为0.791 ± 0.437、1.426 ± 1.104、（162.000 ± 48.046） ng/L、0。佐剂蛋白组各项指标与佐剂组和对照组比较，差异均有统计学意义。提示蛋白疫苗能诱导对沙眼衣原体血清E型株的特异性免疫应答。同源攻击后佐剂蛋白组衣原体持续阳性5周后转阴性，佐剂组及对照组持续阳性。结论 沙眼衣原体rMOMP能刺激恒河猴产生特异性的体液和细胞免疫应答，能诱导恒河猴产生抗沙眼衣原体保护效应。

关键词: 恒河猴

Abstract:

Objective To observe the specific immune responses induced by the recombinant major outer membrane protein （rMOMP） vaccine against Chlamydia trachomatis E serotype in rhesus monkeys. Methods Six rhesus monkeys were equally divided into three groups: adjuvant and protein group vaccinated with purified rMOMP and Freund′s adjuvants, adjuvant group immunized with Freund′s adjuvants only, and control group immunized with phosphate buffer. All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval. Two weeks after the last vaccination, serum, vaginal wash and venous blood samples were collected from the rhesus monkeys, and lymphocytes were isolated from the blood samples. Enzyme linked immunosorbent assay （ELISA） was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA （sIgA） level in wash samples, and methyl thiazolyl tetrazolium （MTT） assay to evaluate the proliferation of lymphocytes after stimulation with Chlamydia trachomatis serotype E elementary bodies. Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies. In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys. Indirect immunofluorescence was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis. Data were statistically analyzed by using one-way analysis of variance and least significant difference （LSD） test with the SPSS 14.0 software. Results The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody （1.718 ± 0.213 vs. 0.841 ± 0.315 and 0.791 ± 0.437, both P < 0.05）, interferon （（1086 ± 121.730） ng/L vs. （409 ± 53.440） ng/L and （162 ± 48.046） ng/L, both P < 0.05）, lymphocyte proliferation index （7.012 ± 1.026 vs. 4.473 ± 1.850 and 1.426 ± 1.104, both P < 0.01） and the diameter of nodus in delayed hypersensitivity assay （（11 ± 2.134） mm vs. （3 ± 0.914） mm and 0, both P < 0.01）. After attack, the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5, and in the other 2 groups from week 1 to 10, but were negative in the adjuvant and protein group from week 6 to 10. Conclusion The rMOMP vaccine can induce a specific, protective, humoral and cellular immune response against Chlamydia trachomatis in rhesus monkeys.

Key words: rhesus monkey