COLLECTINS IN VAGINAL MUCOSAL HOST DEFENSE

阴道粘膜宿主防御中的集合素

• 批准号: 6380027
• 负责人: COLIN MACNEILL
• 金额: $ 23.49万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6380027
• 关键词: candidiasis; clinical research; cytokine; enzyme linked immunosorbent assay; female; hormone regulation /control mechanism; human subject; immunocytochemistry; lectin; mannans; menstrual cycle; mucosa; mucosal immunity; phagocytosis; protein localization; surfactant; tissue /cell culture; vagina; vaginitis; women's health

DESCRIPTION (adapted from applicant's abstract): The vaginal mucosa (VM) is one of the critical interfaces between host and pathogens. Normally, it can tolerate and contain a broad range of microbes and cope with foreign proteins including those present in the ejaculate. Hence, VM is likely to be equipped with an arsenal of defenses similar to that of the other such mucosal interfaces, e.g., lung and gut. This proposal focuses on vaginitis, a condition resulting from a breakdown of defenses against organisms that remain largely localized yet can have major consequences for the host and her reproductive capacity. Vaginitis is responsible for up to 15% of patient visits to women's health practices. The sequelae of VM have great effects on public health, and are implicated in disorders ranging from preterm labor and associated morbidity and mortality of the offspring, to increased susceptibility to sexually transmitted disease and progression of cervical neoplasia. Yet, knowledge of vaginal host defenses has lagged behind that of other mucosal surfaces. Here we propose to follow up on evidence obtained by us that SP-A is expressed in VM and is present in VF by testing the hypotheses that: (1) SP-A in VF, as in lung, contributes to host defenses by modulating cytokine production and facilitating phagocytosis; (2) SP-A expression in VM and function in VF, like other mediators of vaginal immunity, vary with menstrual cycle (i.e., is hormonally regulated); and (3) aberrant SP-A homeostasis contributes to recurrent vulvovaginal candidiasis (RVVC), a prevalent, intractable condition of women of reproductive age that is characterized by episodes of exacerbations of inflammation of the VM due to colonization of the vagina with C. albicans. Parameters relevant to SP-A function will be compared: (1) in healthy women at three stages of the menstrual cycle; (2) in healthy women and RVVC patients; and (3) in RVVC patients during remissions and exacerbations of the disease. In VM, SP-A will be localized by immunocytochemistry and gene-specific message levels will be estimated by reverse transcriptase/polymerase chain reaction (RT/PCR). In VF, SP-A levels will be measured by enzyme-linked immunoabsorbent assay (ELISA), SP-A oligomeric size established by gel filtration and ability of SP-A to stimulate production of proximal cytokines and to facilitate phagocytosis. This will be accomplished using THP-1 macrophages adapted to grow as surrogates for vaginal macrophages in a medium that simulates VF fluid with respect to certain determinants of SP-A function, such as pH, electrolyte and cation concentrations. Immunodepletion will be used to determine the contribution made by SP-A to stimulation of cytokine production by TPH-1 cells.

描述（改编自申请人的摘要）：阴道粘膜（VM）是 宿主和病原体之间的关键界面之一。 正常情况下是可以的 耐受并包含多种微生物并应对外来蛋白质 包括精液中存在的物质。 因此，VM很可能配备 具有与其他粘膜相似的防御手段 界面，例如肺和肠。 该提案的重点是阴道炎， 由于对残留有机体的防御崩溃而导致的状况 很大程度上是本地化的，但可能会对主人和她产生重大影响 生殖能力。 高达 15% 的患者患有阴道炎 参观妇女的健康实践。 VM后遗症影响很大 公共卫生，并与早产和早产等疾病有关 后代的相关发病率和死亡率，以增加 性传播疾病的易感性和宫颈癌的进展 瘤形成。 然而，对阴道宿主防御的了解却落后于人类的了解。 其他粘膜表面。 在此，我们建议对所获得的证据进行跟进 通过检验假设，我们知道 SP-A 在 VM 中表达并且在 VF 中存在 (1) VF 中的 SP-A 与肺中一样，通过调节来促进宿主防御 细胞因子的产生并促进吞噬作用； (2) SP-A在VM中的表达 与阴道免疫的其他介质一样，VF 中的功能和功能也随 月经周期（即受激素调节）； (3) 异常 SP-A 体内平衡导致复发性外阴阴道念珠菌病 (RVVC) 育龄妇女普遍存在的棘手病症是 其特点是 VM 炎症加剧，原因是 白色念珠菌在阴道定植。 SP-A相关参数 功能比较： (1) 健康女性的三个阶段 月经周期； (2)健康女性和RVVC患者；和 (3) 在 RVVC 中 疾病缓解和恶化期间的患者。 在VM中，SP-A将 通过免疫细胞化学和基因特异性信息水平进行定位 通过逆转录酶/聚合酶链反应（RT/PCR）估计。 在VF中， SP-A水平将通过酶联免疫吸附测定（ELISA）测量， 通过凝胶过滤确定 SP-A 寡聚体大小以及 SP-A 的能力 刺激近端细胞因子的产生并促进吞噬作用。 这将使用适应生长的 THP-1 巨噬细胞来完成 在模拟 VF 液体的培养基中替代阴道巨噬细胞 SP-A 功能的某些决定因素，例如 pH、电解质和 阳离子浓度。 免疫耗竭将用于确定 SP-A 对刺激 TPH-1 产生细胞因子的贡献 细胞。