不同铝佐剂明显增强布鲁菌外膜蛋白31(Omp31)的免疫接种效果  

Different aluminum adjuvants significantly enhances the effect of immunization on Brucella Omp31

作　　者：卿蕊 向晴可 刘众齐 肖非 阳帆[2] QING Rui;XIANG Qingke;LIU Zhongqi;XIAO FeiI;YANG Fan(Department of Pathogenic Biology, School of Laboratory Medicine, Shaoyang University, Shaoyang 422600;Department of Basic Medicine, Xiangnan University, Chenzhou 423000, China)

机构地区：[1]邵阳学院医学检验学院病原生物学教研室,湖南邵阳422600 [2]湘南学院基础医学系,湖南郴州423000

出　　处：《细胞与分子免疫学杂志》2018年第2期123-128,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖南省教育厅科学研究项目(16C1438)

摘　　要：目的探讨磷酸铝(AP)和氢氧化铝(AH)佐剂对布鲁菌外膜蛋白31(Omp31)诱导产生体液、细胞免疫反应以及免疫保护作用的影响。方法制备AP佐剂以及AH佐剂,并与纯化的布鲁菌Omp31进行混合吸附,检测吸附率;将AP、AH吸附完成的Omp31蛋白分别于0、2、4周腹腔注射免疫BLAB/c小鼠,同时设立未吸附佐剂的Omp31蛋白免疫组与PBS免疫组作为对照。分别于0、2、4、6周ELISA检测小鼠血清Ig G、Ig G1、Ig G2a水平以及生殖道分泌物分泌型Ig A(s Ig A)水平;于第6周取小鼠脾细胞体外培养,Omp31刺激细胞48 h后,ELISA检测培养细胞上清中γ干扰素(IFN-γ)、白细胞介素10(IL-10)水平,CCK-8法检测小鼠淋巴细胞增殖情况;小鼠于第6周用布鲁菌强毒株进行攻击,并分别于攻击后1周、2周检测小鼠脾脏组织菌体含量。结果 AP、AH佐剂对于不同浓度Omp31吸附率分别可达到70%与85%以上;ELISA结果显示AP组与AH组诱导产生的血清Ig G、Ig G1、Ig G2a以及生殖道s Ig A水平均在末次免疫2周后到峰值,其中AH组高于AP组,且均显著高于对照组;CCK-8实验结果显示AH组诱导的淋巴细胞增殖显著高于AP组,且均高于对照组,AH组脾细胞上清中的IFN-γ水平显著高于AP组,而IL-10水平无显著差异;强毒株16M攻击后2周,AH组小鼠脾脏菌体载量显著低于AP组。结论 AP佐剂以及AH佐剂均能有效增强布鲁菌Omp31的免疫原性及其抗感染保护作用,且AH佐剂效果好于AP佐剂。Objective To investigate the effect of aluminum phosphate （AP） and aluminum hydroxide （AH） as adjuvants on Brucella outer membrane protein 31 （Omp31） in inducing humoral and cellular immune responses and immune protection. Methods AP and AH adjuvants were prepared and separately mixed with Brucella Omp31 protein to measure the adsorption rates. The AP-and AH-absorbed Omp31 protein were intraperitoneally injected into BLAB/c mice at 0, 2, and 4 weeks, and meanwhile, unabsorbed Omp31 protein and PBS were used as controls. The levels of serum IgG, IgG1, IgG2a and genital tract secretion slgA were determined by ELISA at 0, 2, 4 and 6 weeks. Spleen cells were collected for culture at 6 weeks, and the cells were stimulated by Omp31 for 48 hours followed by the analysis of IFN-γ and IL-10 levels in the supernatants by ELISA, and the determination of lymphocyte proliferation by CCK-8 assay. The mice were challenged with Brucella at 6 weeks, and bacterial content in spleen tissue was determined 1 and 2 weeks later. Results AP and AH could absorb over 70% and 85% of the Omp31 protein, respectively, for solutions at all the tested concentrations. ELISA suggested that serum IgG, IgG1, IgG2a and genital tract slgA levels peaked 2 weeks after the last immunization for both AP and AH groups, and antibody level was higher in the AP and AH groups than the control groups, and higher in the AH group than in the AP group. CCK-8 assay showed that the proliferating rate of lymphocytes induced by the AH group was significantlyhigher than that by the AP group, and the AH group also showed significantly higher IFN-γ level in the supernatant than the AP group, but no significant difference in IL-10 level. The AH group had remarkably lower bacterial load in the spleen than the AP group 2 weeks after challenged by Brucella 16M strain immunogenicity and immune protection of the Brucella Omp31 Conclusion Both AP and AH adjuvants effectively enhanced protein, and AH was superior to AP in this respect.

关 键 词：布鲁菌 Omp31 磷酸铝 氢氧化铝 

分 类 号：R392.12[医药卫生—免疫学] R446.62[医药卫生—基础医学]