COLLECTINS IN VAGINAL MUCOSAL HOST DEFENSE

阴道粘膜宿主防御中的集合素

• 批准号: 6198040
• 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 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6198040
• 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。 通常情况下，它可以 容忍并包含广泛的微生物，并科普外来蛋白质 包括那些存在于精液中的物质。 因此，VM可能会配备 与其他粘膜类似的防御武器库 接口，例如，肺和内脏 这项建议的重点是阴道炎， 由于抵抗残留有机体的防御系统崩溃而导致的状况 大部分是局部的，但可能对宿主和她的身体产生重大影响。 生殖能力 阴道炎是负责高达15%的病人 参观妇女保健诊所。 VM的后遗症对 公共卫生，并涉及从早产和 相关的发病率和死亡率的后代，以增加 性传播疾病易感性与宫颈癌进展 肿瘤形成 然而，对阴道宿主防御的了解已经落后于 其他粘膜表面。 在此，我们建议对以下方面获得的证据采取后续行动： 通过检验假设，我们认为SP-A在VM中表达，并存在于VF中 （1）VF中的SP-A与肺中的SP-A一样，通过调节 细胞因子的产生和促进吞噬作用;（2）VM中SP-A的表达 和功能，在VF，像其他介质的阴道免疫，不同的 月经周期（即，受神经调节）;和（3）异常SP-A 内稳态有助于复发性外阴阴道念珠菌病（RVVC）， 育龄妇女普遍存在的顽固性疾病， 其特征是由于以下原因导致VM炎症加剧 阴道内定植C.白色念珠菌。 SP-A相关参数 功能将进行比较：（1）在健康女性的三个阶段， 月经周期;（2）健康女性和RVVC患者;（3）RVVC 在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， 将通过酶联免疫吸附试验（ELISA）测量SP-A水平， 通过凝胶过滤确定的SP-A寡聚体大小和SP-A 刺激近端细胞因子产生并促进吞噬作用。 这将使用THP-1巨噬细胞来实现，所述巨噬细胞适于生长为 阴道巨噬细胞的替代物在模拟VF流体的培养基中， 关于SP-A功能的某些决定因素，如pH、电解质和 阳离子浓度 免疫耗竭将用于确定 SP-A对TPH-1刺激细胞因子产生的贡献 细胞