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

J4 ›› 2010, Vol. 36 ›› Issue (5): 887-890.

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

杨思睿¹, 翟淑波¹, 吴秀丽², 于永利³, 王丽颖²

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

收稿日期:2010-07-22 出版日期:2010-09-28 发布日期:2010-09-28
通讯作者: 王丽颖（Tel:0431-85619369,E-mail:wlying@mail.jlu.edu.cn） E-mail:wlying@mail.jlu.edu.cn
作者简介:杨思睿（1969-）|男|吉林省镇赉县人|副教授|医学博士|主要从事儿科病原学及心血管疾病的研究。

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

YANG Si-Rui¹, ZHAI Chu-Bo¹, WU Xiu-Li², YU Yong-Li³, WANG Li-Ying²

1.Department of Pediatrics,First Hospital,Jilin University,Changchun 130021,China;2.Department of Molecular Biology,School of Basic Medical Sciences,Jilin University|Changchun 130021|China;3.Department of Immunology,School of Basic Medical Sciences,Jilin University|Changchun 130021|China

Received:2010-07-22 Online:2010-09-28 Published:2010-09-28
Supported by:
国家高技术研究发展计划（863计划）资助课题(2002AA214141)

摘要/Abstract

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Ct T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。

关键词: 沙眼衣原体；T细胞表位；热休克蛋白；疫

Abstract:

Abstract: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),14 C57BL/6 female mice were divided into two groups(H-ctm1 immunized mice,naive mice,7 mice 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 72 h following injection. In the experiment of flow cytometric quantitation,21 C57BL/6 female mice were divided into three groups(H-ctm1 immunized mice,HK-EBs immunized mice,PBS-immunized mice,7 mice every group).After immunization,these mice were infected by Ct. 6 d after 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-ctm1 immunized 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）.[JP3]In the FACS experiment,the mean ratio of CD4+ T cells in H-ctm1 group(18.9%±[JP]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-ctm1 g
roup(5.7%±1.9%) was higher than that in HK-EB group(1.5%±0.5%,P<0.01) and control group(0.02%±0.01%,P<0.01). Conclusion DTH play a role in the process of immune response to H-ctm1 immunized mice.HSP65 fusion protein can stimulate the dendritic cells to up-regulate the levels of MHC（class Ⅰ and class　Ⅱ）and improve MOMP-T-epitopes immunogenicity.

Key words: Chlamydia trachomatis；T cell epitopes；heat shock protein；vaccine

中图分类号:

杨思睿, 翟淑波, 吴秀丽, 于永利, 王丽颖. 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[J]. J4, 2010, 36(5): 887-890.

YANG Si-Rui, ZHAI Chu-Bo, WU Xiu-Li, YU Yong-Li, WANG Li-Ying. Protective effect of fusion protein of hsp65-MOMP-T-epitopes on C. trachomatis genital tract infections in mice[J]. J4, 2010, 36(5): 887-890.

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