沙眼衣原体质粒编码蛋白3的致病性及免疫保护性研究
Pathogenicity and Immune Protection of Chlamydia Trachomatis Plasmid-encoded 3
邓晗 1程瑞琴 2陈土地 1宋雅欣 1梁银迎 1李平露 1赵婉星 1马璟玥 1王惠平 1侯淑萍1
作者信息
- 1. 天津医科大学总医院,天津 300052
- 2. 天津河西区瑞普眼科医院,天津 300204
- 折叠
摘要
目的 探索沙眼衣原体(CT)质粒编码蛋白 3(Pgp3)的致病性及免疫保护性.方法 CT L2 构建株(GFP)、野生株(WT)和质粒缺失株(PF)分别感染Hela细胞,蛋白质免疫印迹法(Western blotting)和间接免疫荧光法(IFA)检测Pgp3 蛋白表达水平;L2 GFP、WT和PF菌株经阴道分别感染C3H小鼠,感染后不同时间点经IFA评估生殖道包涵体形成单位(IFU)数量;组氨酸标记的Pgp3(His-Pgp3)体外刺激人输卵管上皮细胞,24 h后经Hoechst 33 528 染色和流式细胞术检测细胞凋亡情况;L2 WT体外感染L929 和L929-Pgp3 细胞,在感染后 30、60 及 80h固定细胞并计算IFU和细胞核数量.L2 GFP和WT菌株经阴道分别感染C3H小鼠,50 d后各组均以L2 WT菌株攻毒,攻毒后不同时间点评估生殖道IFU数量.结果 L2 GFP Pgp3 蛋白表达水平高于L2 WT,L2 PF无Pgp3 蛋白表达;小鼠感染后第3、7、10 和 14 天,L2 GFP感染组IFU数量显著高于L2 WT和L2 PF感染组,且L2 GFP组感染周期最长;Pgp3 体外干预组细胞凋亡率显著低于空白组,L2 WT体外感染L929 和L929-Pgp3 细胞 60h和 80h后,L929-Pgp3 组IFU值显著高于L929 组,且宿主细胞消亡数量显著低于L929 组;动物实验显示L2 GFP组经L2 WT菌株攻毒后下生殖道IFU数量显著低于L2 WT组,且L2 GFP组感染周期显著短于L2 WT组.结论 Pgp3 可抑制宿主细胞凋亡并促进CT在细胞间播散感染;内源性Pgp3 有良好的免疫保护性.
Abstract
Objective In order to investigate pathogenicity and immune protection of Chlamydia trachomatis(CT)plasmid-encodedprotein3(Pgp3).Methods Hela cells were infected with Chlamydia trachomatis L2 green fluorescent protein(GFP),wild type(WT)and plasmid free(PF)strains respectively,and Pgp3 protein expression was detected by Western blotting and indirect immunofluorescence assay(IFA).C3H mice were infected respectively with L2 GFP,WT and PF strains vaginally.After infection,the number of reproductive inclusion body forming unit(IFU)was assessed by IFA at different time points.Human tubal epithelial cells were stimulated with His-Pgp3 in vitro,and Hoechst 33 528 staining as well as flow cytometry were used to detect cell apoptosis after 24 h.L929 and L929-Pgp3 cells were infected by L2 WT strain in vitro.The number of IFU and nuclei were calculated by fixed cells at 30,60 and 80 h after infection,respectively.C3H mice were respectively infected with L2 GFP and WT strains vaginally.After 50 days,each group was challenged with L2 WT strains.After challenged,the number of IFU in lower reproductive tract of mice was evaluated at different time points.Results The expression level of Pgp3 protein in L2 GFP was higher than that in L2 WT,and no Pgp3 protein was expressed in L2 PF.On 3,7,10 and 14 d after infection,the number of IFU in L2 GFP group was significantly higher than that in L2 WT and L2 PF groups,and the L2 GFP group had the longest infection period.The apoptosis rate of Pgp3 intervened cells was significantly lower than that of blank group.After L929 and L929-Pgp3 cells was infected by L2 WT strain for 60 h and 80 h,the number of IFU in L929-Pgp3 group was significantly higher than that in L929 group,and the number of host cell loss was significantly lower than that in L929 group.Animal experiments showed that the number of IFU in the lower reproductive tract of L2 GFP group was significantly lower than that of L2 WT group,and the infection period of L2 GFP group was significantly shorter than that of L2 WT group.Conclusion Pgp3 can inhibit the apoptosis of host cells and promote the spread of CT among cells.Endogenous Pgp3 has good immune protection.
关键词
沙眼衣原体/质粒编码蛋白/3/致病性/免疫保护性Key words
Chlamydia trachomatis/Plasmid-encoded protein 3/Pathogenicity/Immune protection引用本文复制引用
基金项目
国家自然科学基金(31770194)
天津市自然科学基金(18JCYBJC92500)
天津市医学重点学科(专科)建设项目(TJYXZDXK-057B)
出版年
2024
国家科技期刊平台
NSTL
万方数据