中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2013年
8期
610-614
,共5页
吕慧%李静%孙娜%关冰%王红%赵蔚明
呂慧%李靜%孫娜%關冰%王紅%趙蔚明
려혜%리정%손나%관빙%왕홍%조위명
沙眼衣原体%主要外膜蛋白%重组腺病毒%联合免疫
沙眼衣原體%主要外膜蛋白%重組腺病毒%聯閤免疫
사안의원체%주요외막단백%중조선병독%연합면역
Chlamydia trachomatis%Major outer membrane protein%Recombinant adenovirus%Prime-boost immunization
目的探讨E型沙眼衣原体（Ct）主要外膜蛋白（MOMP）DNA疫苗和重组腺病毒联合免疫小鼠诱导的免疫效应。方法构建、纯化重组腺病毒Ad-MOMP及重组真核表达质粒pVAX1-MOMP。设计4种免疫方案，分别为DNA免疫（ DNA组）、重组腺病毒免疫（ Ad组）、DNA初次免疫-重组腺病毒加强免疫（ DNA/Ad组）、重组腺病毒初次免疫-DNA加强免疫（ Ad/DNA组）。末次免疫后2周检测小鼠血清特异IgG、IgG1、IgG2a、IgA抗体，阴道分泌物SIgA抗体及脾淋巴细胞分泌IFN-γ、IL-10水平。结果DNA组诱导较弱免疫应答，未产生SIgA抗体及Th1反应。 Ad组诱导出Th1反应及SIgA抗体，且血清抗体显著高于DNA组。联合免疫均能诱导明显强于单独免疫的黏膜SIgA、血清抗体及Th1反应。 Ad/DNA组的Th1反应强于DNA/Ad组；而DNA/Ad组的血清抗体和黏膜抗体水平强于Ad/DNA组。结论Ad-MOMP能诱导黏膜免疫及Th1细胞免疫应答，DNA/Ad及Ad/DNA联合免疫产生的特异性免疫应答明显强于单独免疫。其中Ad/DNA的Th1反应优势更明显，DNA/Ad的血清抗体和黏膜抗体反应更强。接种顺序会影响联合免疫的强弱及类型，这为Ct疫苗的设计研究提供新的思路和实验依据。
目的探討E型沙眼衣原體（Ct）主要外膜蛋白（MOMP）DNA疫苗和重組腺病毒聯閤免疫小鼠誘導的免疫效應。方法構建、純化重組腺病毒Ad-MOMP及重組真覈錶達質粒pVAX1-MOMP。設計4種免疫方案，分彆為DNA免疫（ DNA組）、重組腺病毒免疫（ Ad組）、DNA初次免疫-重組腺病毒加彊免疫（ DNA/Ad組）、重組腺病毒初次免疫-DNA加彊免疫（ Ad/DNA組）。末次免疫後2週檢測小鼠血清特異IgG、IgG1、IgG2a、IgA抗體，陰道分泌物SIgA抗體及脾淋巴細胞分泌IFN-γ、IL-10水平。結果DNA組誘導較弱免疫應答，未產生SIgA抗體及Th1反應。 Ad組誘導齣Th1反應及SIgA抗體，且血清抗體顯著高于DNA組。聯閤免疫均能誘導明顯彊于單獨免疫的黏膜SIgA、血清抗體及Th1反應。 Ad/DNA組的Th1反應彊于DNA/Ad組；而DNA/Ad組的血清抗體和黏膜抗體水平彊于Ad/DNA組。結論Ad-MOMP能誘導黏膜免疫及Th1細胞免疫應答，DNA/Ad及Ad/DNA聯閤免疫產生的特異性免疫應答明顯彊于單獨免疫。其中Ad/DNA的Th1反應優勢更明顯，DNA/Ad的血清抗體和黏膜抗體反應更彊。接種順序會影響聯閤免疫的彊弱及類型，這為Ct疫苗的設計研究提供新的思路和實驗依據。
목적탐토E형사안의원체（Ct）주요외막단백（MOMP）DNA역묘화중조선병독연합면역소서유도적면역효응。방법구건、순화중조선병독Ad-MOMP급중조진핵표체질립pVAX1-MOMP。설계4충면역방안，분별위DNA면역（ DNA조）、중조선병독면역（ Ad조）、DNA초차면역-중조선병독가강면역（ DNA/Ad조）、중조선병독초차면역-DNA가강면역（ Ad/DNA조）。말차면역후2주검측소서혈청특이IgG、IgG1、IgG2a、IgA항체，음도분비물SIgA항체급비림파세포분비IFN-γ、IL-10수평。결과DNA조유도교약면역응답，미산생SIgA항체급Th1반응。 Ad조유도출Th1반응급SIgA항체，차혈청항체현저고우DNA조。연합면역균능유도명현강우단독면역적점막SIgA、혈청항체급Th1반응。 Ad/DNA조적Th1반응강우DNA/Ad조；이DNA/Ad조적혈청항체화점막항체수평강우Ad/DNA조。결론Ad-MOMP능유도점막면역급Th1세포면역응답，DNA/Ad급Ad/DNA연합면역산생적특이성면역응답명현강우단독면역。기중Ad/DNA적Th1반응우세경명현，DNA/Ad적혈청항체화점막항체반응경강。접충순서회영향연합면역적강약급류형，저위Ct역묘적설계연구제공신적사로화실험의거。
Objective To investigate the specific immune responses induced by DNA vaccine com-bined with recombinant adenovirus carrying major outer membrane protein ( MOMP) gene of Chlamydia trachom-atis (Ct) serovar E.Methods Recombinant eukaryotic expression plasmid pVAX 1-MOMP and recombinant adenovirus Ad-MOMP were constructed and purified .Four immunization strategies were designed including DNA immunization (DNA group), recombinant adenovirus immunization (Ad group), DNA prime-recombinant ade-novirus boost regimen ( DNA/Ad group ) and recombinant adenovirus prime-DNA boost regimen ( Ad/DNA group) .Two weeks after the final immunization , vaginal wash specimens , blood samples and spleens were col-lected from all mice for the evaluation of humoral and cellular immune responses .Results Th1 responses and mucosal responses were not found in DNA group .Ad group induced both Th 1 responses and local mucosal re-sponses , and its IgG and IgA levels were significantly higher than those in DNA group .DNA/Ad group and Ad/DNA group induced stronger Th 1 responses and humoral responses than DNA and Ad group .Serum antibodies and mucosal antibodies in DNA/Ad group were higher than those in Ad/DNA group, but Th1 responses in DNA/Ad group were impaired than those in Ad/DNA group.Conclusion The recombinant adenovirus could induce Th1 responses and genital mucosal responses .Combined immunization of DNA/Ad or Ad/DNA showed enhanced specific immune responses in comparison with the single immunization with DNA or Ad .The differ-ences between the two combination methods were that DNA /Ad group stimulated higher levels of specific anti-body responses , while Ad/DNA group produced stronger Th 1 responses .The study provided a reference for the enhancement and development of vaccines against Chlamydia trachomatis in further studies .
