重组耻垢分枝杆菌制剂预防幽门螺杆菌感染的作用机理的初步探讨  

Mechanism study of recombinant Mycobacteria smegmatis pharmacon in the inmmunization against Helico-bacter pylori

作　　者：向廷秀[1] 谯敏[2] 姜政[2] 黄爱龙[3] 陶小红[2] 王丕龙[2] 赖国旗[4] 

机构地区：[1]重庆医科大学附属第一医院实验研究中心,400016 [2]重庆医科大学附属第一医院消化内科,400016 [3]重庆医科大学感染性疾病分子生物学教育部重点实验室 [4]重庆医科大学实验动物中心

出　　处：《中华微生物学和免疫学杂志》2007年第11期1032-1036,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30371318）;重庆市卫生局重点项目资助[渝卫教（2007）1号文07-1-007]

摘　　要：目的探讨重组耻垢分枝杆菌实验制剂（recombinant Mycobacterium smegmatis，rMs）预防幽门螺杆菌（Helicobacter pylori，Hp）感染的作用机理。方法实验制剂免疫BLAB／c小鼠4周，用Hp攻击后4周，取血清、胃、脾组织，RT-PCR检测小鼠胃黏膜和脾组织的TH1型细胞因子（IFN-γ、IL-2、IL-12）与TH2型细胞因子（IL-4、IL-6、IL-10）；用ELISA检测针对唧的特异性血清IgG、IgG1、IgG2a和IgA水平；用免疫组化方法检测胃黏膜固有层IgA表达；用MTT检测脾淋巴细胞增殖。结果免疫后BALB／c小鼠特异性抗体显示rMs制剂诱导Hp特异性血清IgG、IgA水平都升高，以IgG2a占优势。用Hp抗原刺激后各免疫组小鼠脾淋巴细胞均有明显增殖。免疫组化显示各免疫组小鼠胃黏膜固有层IgA阳性标记腺体数明显高于对照组。RT-PCR证实，跏攻击之前，各免疫组胃和脾组织已出现了IFN-γ、IL-12、IL-2表达而无IL-4、IL-6、IL-10表达。PBS对照组和单纯耻垢分枝杆菌（Ms）组胃黏膜和脾组织各细胞因子均无表达；Hp攻击后，PBS和单纯Ms组胃组织出现以TH1细胞IFN-γ为主的增生，IFN-γ水平显著高于rMs组（P〈0．05）；而3个rMs制剂组脾脏则出现以TH2细胞（IL-4）为主的增生，IL-4水平显著高于对照组（P〈0．05）。提示该制剂的作用机制是诱导TH1，TH2平衡型免疫应答。结论成功地建立了BALB／c小鼠的Hp感染模型，对rMs制荆的免疫保护机理研究结果显示，rMs能产生以TH1细胞和TH2细胞协同作用的保护性免疫应答。Objective To study the immune mechanism of the recombinant Mycobacteria smegmatis （rMs） pharmacon aganist Helicobacter pylori（Hp） in BLAB/c mice. Methods Seventy-five female BLAB/c mice were randomly divided into 5 groups, 4 weeks post-immunization and 4 weeks post-challenge. All mice were sacri-ficed, the blood, gastric and spleen tissue were collected. The cytokine expression of gastric and spleen was analyzed with semiquantitative reverse transcription-PCR. The levels of the Hp specific serum IgG, IgA, IgG1 and IgG2a was detected by ELISA. Multiplication of mouse lympheeytes was detected by MTF, and IgA of gastric tissue was investigated with immunohistochemistry. Results The specific mucosal IgA and specific serum IgG were induced with the administration of rMs, and significant proliferation of spleen lymphocyte was observed in rMs group after the stimulation of Hp antigens. Meanwhile, rich IgA was found in the epithelium and lamina propria of immunized mice with rMs pharmacon as compared with PBS and Ms group mice after 4 weeks challenge. RT-PCR of cy-tokine revealed that every immunity group had relatively high levels of mRNA for IFN-γ, IL-12 and IL-2 but not IL-4, IL-6 and IL-10. After Hp attacked, the levels of IFN-γ, IL-12 and IL-2 mRNA expression of rMs groups were significantly lower than that of the PBS and Ms group（P〈0.05）. While the level of IL-4 mRNA expression of rMs groups were significantly higher than that of the PBS and Ms group （P〈0.05）. There was no significant difference among any rMs group（P〉0.05）. Conclusion TH1 and TH2 protective response can be elicited with rMs immunization.

关 键 词：幽门螺杆菌 重组耻垢分枝杆菌制剂 作用机理 

分 类 号：R686[医药卫生—骨科学]