T. vaginalis viruses as mucosal immunity modifiers with impact on women's health

描述（由申请人提供）：本研究涉及我们对女性生殖道粘膜免疫防御理解的关键空白，以及改善阴道毛滴虫（TV）感染的诊断、治疗和预防的必要性，阴道毛滴虫是美国和全球最常见的非病毒性传播病原体。这种感染（滴虫病）每年影响800 - 1000万美国人，特别是对妇女和儿童造成严重的医疗、经济和社会后果。在育龄妇女和资源匮乏的社区，电视的流行率最高，种族差异巨大（城市社区的非洲裔美国妇女的流行率高出1.5至10倍）。在电视引起的风险和疾病中，不成比例地影响到少数群体的是早产、出生体重低、宫颈癌和艾滋病毒的高风险。治疗由滴虫病引起的艾滋病毒感染的估计终生费用为1.67亿美元。对目前首选药物甲硝唑的耐药性正在上升到惊人的高比率，接受甲硝唑治疗的孕妇早产的风险甚至更高。几乎一半的受艾滋病毒感染的妇女没有症状，而其他妇女则出现严重的生殖器炎症，这是感染艾滋病毒的另一个风险因素。症状差异，缺乏持久免疫力，高复发率和耐药的机制尚不清楚。包括我们在内的几项研究已经证明了在寄生虫的临床分离株中存在TV特异性dsRNA病毒（TVVs），但对其遗传学、毒力、与炎症反应的相关性、耐药性和滴虫病的医学并发症知之甚少。我们最近发表的数据显示，在阴道炎妇女的临床TV分离株中同时存在多达4种不同的TVV。此外，我们已经证明，在生殖器上皮细胞中，携带一个或多个TVV的TV株诱导增强的病毒依赖性炎症反应。我们提出了一个新的假设，涉及TVVs作为TV毒力的修饰剂和TV感染的症状差异和当前治疗的副作用的因素，从而成为预后/诊断生物标志物和新疗法的关键目标。这一假设将通过以下具体目标来解决：（1）发现TVV-TV-人相互作用的新的分子决定因素，其对阴道上皮免疫功能有影响（2）定义TVV的遗传、生物化学和生物物理特征，其对TV毒力和阴道免疫功能有影响，可用于诊断或治疗方法;（3）收集和检查临床上定义的原发性TV分离株，以将不同的TVV基因型和分子特征与最易患TV可归因疾病的美国人群中的耐药性和症状性滴虫病联系起来。我们的跨学科方法包括生理体外模型系统，新的分子生物学技术和等基因工具，以及寻求治疗滴虫病的妇女的调查。拟议的转化研究将产生对妇女和儿童健康具有高度影响的阴道免疫和感染风险因素的新的基础知识。

DESCRIPTION (provided by applicant): This study deals with critical gaps in our understanding of the mucosal immune defenses of the female genital tract and the need to improve diagnosis, cure, and prevention of infection by Trichomonas vaginalis (TV), which is the most common nonviral sexually transmitted pathogen in the US and worldwide. The infection (trichomoniasis) affects 8-10 million Americans each year with serious medical, economic, and social consequences especially for women and children. The prevalence of TV is highest in women of reproductive age and in low-resource communities with an enormous racial disparity (1.5 to 10 times more prevalent in African-American women from urban communities). Among TV-attributable risks and pathologies disproportionately affecting minority populations are pre-term birth, low birth weight, cervical cancer, and higher risk of HIV. The estimated lifetime cost of treating trichomoniasis-attributable HIV infections is $167 million. Resistance to the current drug of choice, metronidazole, is rising to alarmingly high rates, and pregnant women treated with metronidazole are at even higher risk of pre-term birth. Almost half of TV-infected women are asymptomatic while the others develop a severe genital inflammation, which is an additional risk factor for HIV acquisition. The mechanisms of symptom disparity, lack of lasting immunity, high recurrence rate, and resistance to treatment are unknown. Several studies including ours have documented the presence of TV-specific dsRNA viruses (TVVs) in clinical isolates of the parasite, but little is known about their genetics, virulence, relevance to the inflammatory reaction, drug resistance, and medical complications of trichomoniasis. Our recently published data show the simultaneous presence of as many as 4 distinct species of TVVs in clinical TV isolates from women with vaginitis. Furthermore, we have demonstrated that in genital epithelial cells, TV strains harboring one or more TVVs induce a heightened, virus- dependent inflammatory response. We propose a novel hypothesis that implicates TVVs as modifiers of TV virulence and as factors underlying symptom disparity of TV infections and current therapy's side effects, thus emerging as key targets for prognostic/diagnostic biomarkers and new therapies. This hypothesis will be addressed by the following Specific Aims: (1) discover novel molecular determinants of TVV-TV-human interactions with impact on vaginal epithelial immune function (2) define genetic, biochemical, and biophysical characteristics of TVVs with impact on TV virulence and vaginal immune function that could be used for diagnostic or therapeutic approaches; (3) collect and examine clinically defined primary TV isolates to link distinct TVV genotypes and molecular characteristics to drug resistance and symptomatic trichomoniasis in a US population most vulnerable to TV-attributable disease. Our interdisciplinary approach includes a physiological in-vitro model system, novel molecular biology techniques and isogenic tools, as well as a survey of women seeking treatment for trichomoniasis. The proposed translational research will generate new basic knowledge of vaginal immunity and infection risk factors with high impact on women's and children's health.