EVALUATION OF C31G VS NONOXYNOL-9 AS TOPICAL MICROBICIDES

生殖器沙眼衣原体D-K血清型与流行的关系 在美国，性传播感染影响了400多万男性， 妇女和婴儿每年。在女性中，如果不接受治疗，衣原体可能会 从宫颈管内扩散到子宫内膜 (子宫内膜炎)和输卵管(输卵管炎)-- 它被定义为盆腔炎。由于输卵管 疤痕形成和/或卵子运输受损，异位妊娠和 可能会发生不孕不育。显然，衣原体疾病构成了重要的 初级、二级和三级保健问题，妇女在这些问题上承担 特别的负担，因为她们生育不良的风险增加了 后果。迫切需要有效的局部杀微生物剂来帮助 预防和控制性传播感染。 这笔赠款的目的是评估专题论坛的行动 杀菌剂C316(及其衍生物B58A和B64D)与壬醇-9的比较 PH 5.7和7.0对沙眼衣原体E亚型及其靶宿主细胞的影响。 杀微生物剂的浓度将被量化为 极化的人生殖器上皮细胞的相容性和细胞毒性 使用二乙酸荧光素和丙二酸丙酯的细胞(鳞状、柱状) 在特定目的1中添加碘，在特定目的中使用分离的衣原体。 放射性标记的传染性基本体(EB)和代谢 活性网状小体将通过SDS-PAGE分析抗原的丢失， 免疫印迹、放射自显影和免疫电子显微镜 生存能力：暴露于杀微生物剂的EB也将被检测附着到 宿主上皮细胞与包涵体发育。在具体目标3中， 杀菌剂暴露对极化的人生殖器上皮细胞的影响 (A)正常感染和(B)持续感染衣原体 经荧光和透射电子显微镜检查，以及 雌激素和雌激素加孕酮对这种作用的调节作用将是 特定目标评估4.未暴露和暴露于杀微生物剂的感染 极化的人类上皮细胞将与早产儿增加进行比较 衣原体抗原从包涵体中释放并分泌到宿主 细胞表面包埋后免疫标记的电子显微镜观察 在Lowicryl中加工的样品(具体目标5)。最后，我们将 确定炎症反应细胞是否迁移到衣原体- 受感染的极化上皮细胞通过接触杀菌剂进行调节。

Genital Chlamydia trachomatis serovars D-K are responsible for epidemic sexually transmitted infections in the USA, affecting over 4 million men, women and infants per year. In women, without treatment, chlamydiae may spread canalicularly from the endocervical canal to the endometrium (endometritis), and the fallopian tubes (salpingitis)- the constellation of which is defined as pelvic inflammatory disease. As a result of tubal scarring and/or impaired ovum transportation, ectopic pregnancy and infertility can occur. Clearly, chlamydial diseases constitute significant primary, secondary and tertiary health care concerns in which women bear a special burden because of their increased risk of adverse reproductive consequences. Effective topical microbicides are urgently needed to help prevent and control sexually transmitted infections. The purpose of this grant is to evaluate the action of the topical microbicide C316 (and its derivatives B58A and B64D) versus nonoxynol-9 at pH 5.7 and 7.0 on Chlamydia trachomatis serovar E and its target host cell. The concentrations of the microbicides will be quantitated for compatibility and cytotoxicity with polarized human genital epithelial cells (squamocolumnar, columnar) using fluorescein diacetate and propidium iodide in Specific Aim 1 and with isolated chlamydiae in Specific Aim 2. Both radiolabeled infectious elementary bodies (EB) and metabolically active reticulate bodies will be analyzed for loss of antigens by SDS-PAGE, Western blots, autoradiography, and immunoelectron microscopy and for viability: microbicide-exposed EB will also be assayed for attachment to host epithelial cells and inclusion development. In Specific Aim 3, the effect of microbicide exposure on polarized human genital epithelial cells (a) normally infected and (b) persistently infected with chlamydiae will be examined by fluorescence and transmission electron microscopy, and modulation of the effect by estrogen and estrogen plus progesterone will be assessed in Specific Aim 4. Unexposed and microbicide-exposed infected polarized human epithelial cells will be compared for increased premature chlamydial antigen release from inclusions and antigen secretion to host cell surface - by post-embedding immunolabeling of electron microscopy samples processed in Lowicryl (Specific Aim 5). Finally, we shall determine wether or not inflammatory response cell migration to chlamydiae- infected polarized epithelial cells is modulated by microbicide exposure.