摘要
目的 探讨Toll样受体2（TLR2）和TLR4在小鼠沙眼衣原体生殖道感染中的作用。方法用1×104包涵体形成单位的MoPn沙眼衣原体小鼠肺炎株经生殖道感染野生型（WT）小鼠（n=11）、 TLR2基因缺陷 （TLR2-/-）小鼠（n=14）和TLR4-/-小鼠（n=11）, 复制生殖道沙眼衣原体感染模型。在感染后第3、7、10、17、24、31、38、45天取阴道棉拭子, 免疫荧光法检测衣原体包涵体数量, ELISA检测白细胞介素1α（IL-1α）、IL-6和巨噬细胞炎性蛋白2（MIP-2）水平。感染后第70天, 采用免疫荧光法分析血清抗体类型和效价; 用MoPn沙眼衣原体感染无菌分离的腹腔巨噬细胞, 培养24 h, ELISA测定上清液中IL-1α、IL-6和MIP-2含量; 体外用紫外线灭活的衣原体抗原刺激无菌分离的脾细胞, 培养72 h, ELISA测定上清液中γ干扰素（IFN-γ）、IL-17、IL-4和IL-5水平。结果 与WT小鼠相比, TLR2-/-或TLR4-/-小鼠在各时间点的生殖道带菌量无差异、且带菌持续时间相同, 感染后第38天, 所有小鼠均清除了下生殖道感染的衣原体。TLR2-/-小鼠巨噬细胞产生的IL-1α、IL-6和MIP-2水平均明显低于WT小鼠,而TLR4-/-小鼠巨噬细胞产生的3种炎症细胞因子水平均与WT小鼠无显著性差异; TLR2-/-小鼠棉拭子标本具有较低水平的炎症细胞因子。所有小鼠脾细胞均产生高水平的IFN-γ和IL-17、 较低水平的IL-4和IL-5、血清IgG2a/ IgG1比值均大于1, 但各组间无显著性差异。结论 TLR2而不是TLR4介导了沙眼衣原体生殖道感染的早期免疫应答, 但沙眼衣原体诱导的适应性免疫应答可能既不依赖于TLR4, 也不依赖于TLR2。
Objective To evaluate the roles of Toll-like receptor 2 （TLR2） and TLR4 in immune responses to Chlamydia trachomatis （Ct） genital tract infection in mice. Methods The wild type （ WT, n = 11 ）, TLR2 -/- （ n = 14） and TLR4 -/- （ n = 11 ） mice were inoculated intravaginally with 1 x 104 inclusion forming units （IFUs） of live C. muridarum （strain MoPn/Nigg） to establish the models of Ct genital tract infection. The vaginal swabs were taken at the 3rd, 7th, 10th, 11th, 24th, 31 st, 38th, 45th day after the infection. Immunofluorescence assay was used to quantity live organisms from each swab. The inflammatory cytokines interleukin-1α （IL-1α）, IL-6 and macrophage inflammatory protein 2 （MIP-2） were measured using ELISA. The ？Oth day post infection, the titers of mouse serum Ct-specific antibody isotypes were determined using an immunofluorescence technique; after the macrophages harvested from peritoneal cavity were infected with MoPn strain for 24 hours, the culture supernatants were examined for the contents of cytokines IL-1α, IL-6 and MIP-2 using ELISA. The harvested splenocytes were stimulated with UV-inactivated chlamydial antigens for 72 hours, and the culture supernatants were measured for the levels of cytokines interferon γ （IFN-γ）, IL-17, IL-4 and IL-5 using ELISA. Results No significant difference was observed in the number of IFUs from the vaginal swabs at any time points post infection among WT,TLR2-/- and TLR4-/- mice. All mice displayed similar time course of live organism shedding. Until the 38th day, all mice cleared live organism. Macrophages lacking TLR2 produced significantly decreased amounts of IL-1α, IL-6 and MIP-2 compared with those in WT mice; however, there was no significant difference between WT and TLR4-/- mice. The vaginal swab samples from TLR2 -/- mice also had lower levels of inflammatory cytokines. Splenocytes from the three groups of mice all produced high levels of both IFN-α, and IL-17, and low levels of both IL-4 and IL-5, and no significant difference was found among the three groups. The ratio of serum IgG2a/IgG1 in the three groups of mice was greater than 1, and no significant difference was found among the three groups. Conclusion TLR2, rather than TLR4 mediates early immune response following Ct genital tract infection in mice. However, neither TLR4 nor TLR2 is required for adaptive immune responses induced by Ct genital tract infection.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第10期1336-1341,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
河北北方学院创新人才培育项目(CXRC1316)
美国国立卫生研究院基金(IR01 AI47997-01)
