Differential expression of mRNA encoding cytokines and chemokines in the reproductive tract after infection of mice with Chlamydia trachomatis. Mus musculus

Chlamydia trachomatis is an obligate intracellular pathogen and the most frequently reported sexually transmitted bacteria in the United States. Infection with Chlamydia trachomatis targets epithelial cells within the genital tract which respond by secreting chemokines and cytokines. Persistent inflammation can lead to fibrosis, tubal infertility and/or ectopic pregnancy. Our objective was to determine the inflammatory mediators involved in clearance of low-grade infection and the potential involvement in chronic inflammation. Our hypothesis was that mRNA encoding pro-inflammatory cytokines and chemokines will be differentially expressed in the female reproductive tract of mice infected with C. trachomatis at both 28 and 35 days post-infection compared to controls. Superarray analysis was performed using RT2 Profiler PCR arrays for mouse Cytokines and Chemokines (Qiagen). Overall design: Six to eight week old female C3H/HeJ mice (Jackson Laboratories, Sacramento, CA) were pretreated with 2.5 mg medroxyprogesterone acetate (SICOR Pharmaceuticals) on Days -10 and -3 before infection. On Day 0, mice were infected via vaginal challenge with 0 or 5 x 102 inclusion-forming units (IFUs) of C. trachomatis, serovar D in 0.01 mL of Eagle Minimal essential media (MEM, Gibco). Mice were killed on day 28 (n = 3 for control and infected) in experiment 1 and day 35 (n = 3 for control and infected) in experiment 2. Vaginal tissue, uterine horns and the oviducts were collected and processed for superarray analysis.

沙眼衣原体（Chlamydia trachomatis）是一种专性胞内病原体，也是美国报告病例数最多的性传播细菌。该病原体感染会靶向生殖道上皮细胞，宿主上皮细胞会通过分泌趋化因子与细胞因子产生应答。持续炎症反应可引发纤维化、输卵管性不孕及异位妊娠。本研究旨在明确参与低度感染清除的炎症介质，以及其在慢性炎症发生中的潜在作用。研究假设为：相较于对照组，感染沙眼衣原体（C. trachomatis）的小鼠雌性生殖道中，编码促炎细胞因子与趋化因子的mRNA在感染后28天及35天均会呈现差异表达。实验采用Qiagen公司的小鼠细胞因子与趋化因子RT2 Profiler PCR芯片（RT2 Profiler PCR Arrays）进行超阵列分析。实验整体设计如下：将6至8周龄的雌性C3H/HeJ小鼠（购自加利福尼亚州萨克拉门托市杰克逊实验室（Jackson Laboratories））于感染前第-10天和第-3天，预先给予2.5mg醋酸甲羟孕酮（购自SICOR制药公司）。感染第0天，小鼠经阴道接种0或5×10²个包涵体形成单位（IFUs）的沙眼衣原体血清型D，接种体积为0.01mL的伊格尔最低必需培养基（MEM, Gibco）。实验1于感染后第28天处死小鼠（对照组与感染组各n=3），实验2于感染后第35天处死小鼠（对照组与感染组各n=3）。收集小鼠阴道组织、子宫角及输卵管，处理后用于超阵列分析。