EVALUATION OF C31G VS NONOXYNOL-9 AS TOPICAL MICROBICIDES

C31G 与 NONOXYNOL-9 作为外用杀菌剂的评价

• 批准号: 6099894
• 负责人: Priscilla B. Wyrick
• 金额: $ 10.86万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099894
• 关键词: Chlamydia trachomatis; antiinfective agents; bacterial antigens; basolateral membrane; cell migration; cellular polarity; chronic disease /disorder; drug screening /evaluation; epithelium; female; human tissue; inflammation; nonoxynol 9; progesterone; topical drug application

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.

生殖器沙眼衣原体 D-K 血清型是流行的罪魁祸首 在美国，性传播感染影响了超过 400 万男性， 每年妇女和婴儿。在女性中，如果不进行治疗，衣原体可能会感染 从子宫颈管呈小管状扩散至子宫内膜 （子宫内膜炎）和输卵管（输卵管炎） - 星座 这被定义为盆腔炎。由于输卵管 疤痕和/或卵子运输受损、宫外孕和 可能会发生不孕症。显然，衣原体疾病对 妇女承担的初级、二级和三级卫生保健问题 由于不良生殖风险增加而造成特殊负担 结果。迫切需要有效的局部杀菌剂来帮助 预防和控制性传播感染。 这笔赠款的目的是评估主题的行动 杀菌剂 C316（及其衍生物 B58A 和 B64D）与 nonoxynol-9 在 沙眼衣原体血清型 E 及其靶宿主细胞的 pH 5.7 和 7.0。 杀微生物剂的浓度将被定量 与极化人类生殖上皮的相容性和细胞毒性 使用荧光素二乙酸酯和丙锭的细胞（鳞柱状、柱状） 具体目标 1 中的碘化物和具体目标 2 中的分离衣原体。 放射性标记的传染性基本体（EB）和代谢性基本体 将通过 SDS-PAGE 分析活性网状体的抗原损失， 蛋白质印迹、放射自显影和免疫电子显微镜 活力：还将检测接触过杀菌剂的 EB 的附着情况 宿主上皮细胞和包涵体发育。在具体目标 3 中， 杀菌剂暴露对极化的人类生殖器上皮细胞的影响 (a) 正常感染和 (b) 持续感染衣原体将 通过荧光和透射电子显微镜检查，以及 雌激素和雌激素加孕激素对效果的调节将是 在具体目标 4 中进行评估。未暴露和暴露于杀菌剂的感染者 极化的人类上皮细胞将被比较以增加早产 衣原体抗原从包涵体中释放并分泌至宿主 细胞表面 - 通过电子显微镜的包埋后免疫标记 Lowicryl 处理的样品（具体目标 5）。最后，我们将 确定炎症反应细胞是否迁移至衣原体- 受感染的极化上皮细胞受到杀微生物剂暴露的调节。