E型沙眼衣原体重组主要外膜蛋白诱导出的小鼠免疫效应

目的 检验自行研制的E型沙眼衣原体重组主要外膜蛋白(rMOMP)诱导小鼠产生的特异性免疫应答效应.方法 3～4周清洁级BALB/C雌鼠36只,分为佐剂组、单独组和对照组,每组12只.佐剂组采用纯化的rMOMP 50 μg和等体积的弗氏佐剂、单独组单用纯化的rMOMP 50μg、对照组单用PBS200μL,于第0、2、4周分别双侧股四头肌进行免疫.ELISA法检测小鼠血清中沙眼衣原体特异性IgG抗体以及阴道冲洗液中沙眼衣原体特异性sIgA抗体,ELISA法检测细胞因子IFN-γ,MTT法检测小鼠脾淋巴细胞特异性增殖反应;同源攻击后阴道宫颈脱落细胞沙眼衣原体培养,观察小鼠的迟发型超敏反应以及小鼠的血清抗体中和试验.结果 佐剂组小鼠血清沙眼衣原体特异性IgG抗体A_(405)值为0.641±0.059,淋巴细胞增殖指数5.085±1.291,迟发型超敏反应右足足垫增厚0.324 ±0.054 mm.单独组小鼠血清特异性IgG抗体A_(405)值为0.424±0.015,淋巴细胞增殖指数3.123±0.840,迟发型超敏反应右足足垫增厚0.272±0.064mm.佐剂组各项指标的检测结果都高于单独组(P<0.05)和对照组(P<0.01).结论 沙眼衣原体rMOMP能刺激机体产生有效的特异性体液和细胞免疫.

Immune responses induced by recombinant major outer membrane protein of Chlamydia trachomatis serovar E

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.