沙眼衣原体T细胞表位融合蛋白对沙眼衣原体感染小鼠的保护机制  

Protective effect of fusion protein of hsp65-MOMP-T-epitopes on C.trachomatis genital tract infections in mice

作　　者：杨思睿[1] 翟淑波[1] 吴秀丽[2] 于永利[3] 王丽颖[2] 

机构地区：[1]吉林大学第一医院儿科,吉林长春130021 [2]吉林大学基础医学院分子生物学教研室,吉林长春130021 [3]吉林大学基础医学院免疫学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2010年第5期887-890,共4页Journal of Jilin University：Medicine Edition

基　　金：国家高技术研究发展计划(863计划)资助课题(2002AA214141)

摘　　要：目的:探讨沙眼衣原体(Ct)感染的T细胞表位融合蛋白疫苗(H-ctm1)对小鼠生殖道感染保护作用的机制,为研制和开发CtT细胞表位融合蛋白疫苗奠定理论和实验基础。方法:建立阴道感染沙眼衣原体的动物模型。迟发型超敏反应(DTH)实验中,将14只小鼠随机分2个实验组:H-ctm1免疫组和未免疫组(每组7只)。测量每只小鼠双侧后脚掌厚度,左后脚掌足底皮下注射蔗糖-磷酸盐运送液(SPG),右后脚掌皮下注射热灭活EB(HK-EBs),每组分别比较左右后脚掌足底厚度和足底注射前后的厚度;在流式细胞术分析实验中,将21只雌性小鼠随机分为3组(每组7只),分别为H-ctm1免疫组、HK-EBs免疫组(阳性对照)和磷酸盐缓冲液(PBS)免疫组(阴性对照)。免疫后在各组小鼠阴道内接种Ct。在阴道感染Ct后第6天取外周血对CD4+和CD8+T细胞进行流式细胞术分析。结果:DTH实验中,未免疫组小鼠左或右后脚掌注射前后足底厚度比较差异无显著性(P>0.05),注射前或注射后左右后脚掌足底厚度比较差异无显著性(P>0.05)。免疫组小鼠注射前左右后脚掌足底厚度比较差异无显著性(P>0.05),注射后左后脚掌足底厚度高于右后脚掌(P<0.05)。左后脚掌注射前后足底厚度比较差异无显著性(P>0.05),右后脚掌注射后足底厚度高于注射前(P<0.05)。流式细胞术记数法测3组活化的CD4+和CD8+T细胞,H-ctm1组CD4+和CD8+T细胞阳性率均高于HK-EBs组(P<0.05或P<0.01);H-ctm1组和HK-EBs组CD4+和CD8+T细胞阳性率均高于PBS组(P<0.01)。结论:DTH参与了H-ctm1免疫小鼠的免疫应答;在H-ctm1中,HSP65的存在刺激DC细胞上调MHC(Ⅰ类途径和Ⅱ类途径)分子的表达水平,提高沙眼衣原体主要外膜蛋白表位的免疫原性。Objective To study the mechanism of fusion protein of hsp65-MOMP-T-epitopes （H-ctm1）against C.trachomatis （Ct）genital tract infections that will underlay further research for T-epitopes Ct vaccine. Methods Models for Ct genital tract infections of mice were established.In the experiment of delayed type hypersensitivity（DTH）,14C57BL/6female mice were divided into two groups（H-ctm1immunized mice,naive mice,7mice every group）.The thickness of bilateral hind footpad of every mouse was measured,then,the left hind footpad was injected with SPG,and the right hind footpad with heat-killed chlamydia （HK-EBs）.The thickness of hind footpad was remeasured at 72hfollowing injection.In the experiment of flow cytometric quantitation,21 C57BL/6female mice were divided into three groups （H-ctm1immunized mice,HK-EBs immunized mice,PBS-immunized mice,7mice every group）.After immunization,these mice were infected by Ct. 6dafter infection,the CD4＋ and CD8＋ T cells were dectected with flow cytometry （FACS）.Results For the experiment of DTH,in the group of naive mice,there was no significant difference in thickness between pre-injection and post-injection whatever right or left hind footpad （P〉0.05）,also between right and left hind footpad whatever pre-injection and post-injection （P〉0.05）.In the group of H-ctm1immunized mice,there was no significant difference in thickness between right and left hind footpad before injection （P〉0.05）,however,the difference was significant after injection （P〈0.05）.For left hind footpad,the difference was not significant between before and after injection （P〉0.05）;for right hind footpad,the difference was significant between before and after injection （P〈0.05）.In the FACS experiment,the mean ratio of CD4＋ T cells in H-ctm1group （18.9%± 2.2%）was higher than that in HK-EB group （11.3%±3.1%,P〈0.05））and control group （0.05%±0.07%,P〈0.01）.And the mean ratio of CD8＋cells in H-ctm1group （5.7%±1.9%）was higher than

关 键 词：沙眼衣原体 T细胞表位 热休克蛋白 疫苗 

分 类 号：R374.1[医药卫生—病原生物学] R-332[医药卫生—基础医学]