沙眼衣原体pORF5质粒蛋白诱发小鼠生殖道免疫损伤初步研究  

Prefinfnmry study on gemtal tract immune injury in mice induced by pORt＇5 plasmid protein of Cidamydia traclmmatis

作　　者：邓红玉[1] 李忠玉[1] 吴移谋[1] 周辉[1] 马康康[1] 陆春雪[1] 钟光明[2] 

机构地区：[1]南华大学医学院微生物学与免疫学教研室,衡阳421001 [2]美国德州大学圣安东尼奥健康科学中心

出　　处：《中华微生物学和免疫学杂志》2013年第2期107-111,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30970165、81102230）;渤南省高校科技创新团队（湘教通[2010]212号）;湖南省普通高等学校重点实验室资助项目

摘　　要：目的研究沙眼衣原体（Ch／amyd／atrachomatis，ct）pORt5质粒蛋白对小鼠生殖道组织的免疫损伤作用，并初步探讨其损伤机制。方法表达纯化GST-pORF5融合蛋白，融合蛋白经PreScissionProtease酶切和去内毒素处理后分别在第1、3、6天接种于BALB／c小鼠阴道后穹窿，第7天处死小鼠，分离小鼠生殖道组织，肉眼大体观察并进行炎症病理评分；ELISA方法检测血清、阴道灌洗液和小鼠脾细胞培养上清中的TNF-a、IL-1β和IL石细胞因子水平。结果pORF5蛋白接种组小鼠输卵管壶腹部与峡部出现不同程度的肿胀，周围结缔组织发生黏连，并出现了不同程度的扭曲与积水，而PBS和csr（谷胱甘肽S-转移酶）蛋白对照组输卵管组织未出现明显改变；同时生殖道组织炎症病理积分明显高于PBS对照组（P〈0．01）和GST对照组（P〈0．01）；pOaF5蛋白接种组小鼠脾细胞培养上清、阴道分泌物中TNF-a、IL-IB、Ib6水平均显著高于PBS和GST蛋白对照组（P〈0．05）；血清中TNF-a、IL-1B水平显著高于PBS和GST蛋白对照组（P〈0．01）。结论pOaF5蛋白能引起BAIB/c小鼠生殖道组织免疫损伤，该损伤机制可能与TNF-a、IL-115、IL石等炎症因子水平升高有关。Objective To investigate the immlme injury in genital tract of BALB/c mice induced by plasmid protein pORF5 of Gh/amyd/a trachoma and its possible mechanism. Methods GST（glutathi- one-S-tmnsferases） -pORF5 fusion protein was expressed and digested with PreScission Protease to obtain the target protein without C, ST tag. After further purification and endotoxm removal, pORF5 protein was injected into the posterior foroix of BALB/c mice on day 1, 3 and 6, while the control groups were injected with PBS or GST protein respectively, and then all the mice were sacrificed on day 7 to evaluate genital tract gross pa- thology and histopathological characterization. The levels of TNF-ct, IL-1β and IL-6 in serum, splenocytes culture supernatant and vaginal douche were detected by ELISA. Results Mice in pORF5 group developed different degrees of swelling in isthmic portion and ampulla of uterine tube, connective tissue adhesion and hydrosalpinx in the genital tract tissues, while the PBS group and the GST group did not show any obvious change. The inflammatory score showed that the genital tract pathology in pORF5 group was much mare se- vere than PBS and GST control groups （P〈0.01）. The levels of TNF-ot, IL-113 and IL-6 in vaginal douche and splenecytes culture supematants in pORF5 group were obviously higher than those d PBS and GST groups （P〈0.05）. The levels of TNF-a and IL-l[3 in serum were also higher than those d GST and PBS groups （ P〈0.01 ）. Conclusion pORF5 plasmid protein could induce pathological immune response in the genital tract of BALB/c mice, which may be associated with the increase of the production of the idlammato- ry cytokines TNF-a, 1L-1β and IL-6 in BALB/c mice.

关 键 词：沙眼衣原体 pORF5质粒蛋白 免疫损伤 细胞因子 

分 类 号：R392.1[医药卫生—免疫学]