Genital Immune, Mucosal, and Viral Effects of Female Genital Schistosomiasis in Tanzania

项目总结/摘要 女性生殖器血吸虫病（FGS），由寄生虫血吸虫引起，影响40 在非洲的数百万女孩和妇女。寄生虫卵通过粘膜组织迁移，诱导宿主免疫 导致女性生殖道糜烂和粘膜破裂的反应，症状包括 生殖器分泌物、出血、疼痛和不孕。慢性生殖道损伤和症状持续后， 吡喹酮治疗约70%的妇女，即使吡喹酮有效地杀死寄生蠕虫， 在尿中排泄卵，并解决膀胱中的大多数组织病理学。相反，寄生虫卵 在治疗后被困在生殖器组织中，根据尸检研究，已知它们会诱导粘膜 以肉芽肿形成和纤维化为特征的免疫反应。FGS是一种被忽视的热带疾病， 在我们对其细胞和分子病理生理学的理解方面存在重要的知识缺口。我们 不知道免疫细胞对S.生殖器组织中的血吸虫卵， FGS对上皮细胞屏障的影响，以及FGS相关的细胞和分子变化是否增加 易受病毒性生殖道感染。提出这一建议的理由是， 缺口可能导致靶向免疫调节、组织修复或病毒抑制干预， 生殖器粘膜受损。根据我们的初步数据，我们的中心假设是S。血吸虫卵 调节宫颈免疫力，减少抗病毒免疫细胞， 上皮屏障，并增加HSV-2的复发，导致发病率和持续的症状， 甚至在吡喹酮治疗后也有FGS。为了验证这一假设，我们将研究90名患有S。吸虫 90例对照组无感染。女人与S。将在基线时接受吡喹酮治疗 随访12个月内，如果持续或复发S。检测到嗜血杆菌。我们将追捕三名 具体目的：1）明确S.血吸虫感染，之前和 吡喹酮后; 2）确定与生殖器上皮完整性破坏相关的分子机制， 患有S. 3）定量S.埃及血吸虫感染 生殖器HSV-2再激活的频率、强度和持续时间。在第一个目标中，我们将收集宫颈细胞， 通过流式细胞术和质谱细胞术对细胞进行刷洗和表征。第二个目标是分离上皮细胞 通过宫颈刷收集并进行RNA-Seq，以阐明上皮细胞特异性的基因和途径， 完整在第三个目标中，我们将在一个月内量化来自队列的女性的HSV-2病毒脱落， HSV-2血清阳性（n=90）。在探索性分析中，我们还将通过以下方法检查阴道病毒组： 宏基因组测序这项拟议中的研究意义重大，因为它可能会发现新的治疗方法， 为数百万患有FGS的女孩和妇女提供帮助。此外，它还推进了寄生虫和病毒的新研究， 扩大我们对蠕虫、粘膜免疫和病毒感染之间相互作用的理解。

Project Summary/Abstract Female genital schistosomiasis (FGS), caused by the parasitic worm Schistosoma haematobium, affects 40 million girls and women in Africa. Parasite eggs migrate through mucosal tissue, inducing a host immune reaction that leads to erosions and mucosal breaches of the female genital tract with symptoms including genital discharge, bleeding, pain, and infertility. Chronic genital tract damage and symptoms persist after praziquantel therapy in ~70% of women, even though praziquantel effectively kills parasite worms, reduces excretion of eggs in urine, and resolves most tissue pathology in the bladder. In contrast, parasite eggs remain trapped in genital tissue post-treatment where, from autopsy studies, they are known to induce a mucosal immune response characterized by granuloma formation and fibrosis. FGS is a neglected tropical disease and there are important knowledge gaps in our understanding of its cellular and molecular pathophysiology. We do not know the profiles or functions of immune cells that respond to S. haematobium eggs in genital tissue, the effects of FGS on the epithelial cell barrier, and if FGS-related cellular and molecular changes increase susceptibility to viral genital tract infections. The rationale for this proposal is that addressing these knowledge gaps could lead to targeted immunomodulatory, tissue reparative, or viral suppressive interventions to restore damaged genital mucosa. Based on our preliminary data, our central hypothesis is that S. haematobium eggs in the genital mucosa modulate cervical immunity and decrease anti-viral immune cells, cause breakdowns in the epithelial barrier, and increase recurrences of HSV-2, resulting in the morbidity and persistent symptoms of FGS even after praziquantel therapy. To test this hypothesis, we will study 90 women with S. haematobium infection and 90 controls without. Women with S. haematobium will receive praziquantel treatment at baseline and during 12 months of follow up if persistent or recurrent S. haematobium is detected. We will pursue three specific aims: 1) Define the genital mucosal immune cell composition in S. haematobium infection, before and after praziquantel; 2) Determine the molecular mechanisms linked to breakdown of genital epithelial integrity in women with S. haematobium infection; and 3) Quantify the effect of S. haematobium infection on the frequency, intensity, and duration of genital HSV-2 reactivation. In the first aim, we will collect cervical cells by brush and characterize cells by flow and mass cytometry. In the second aim, we will isolate epithelial cells collected by cervical brush and perform RNA-Seq to elucidate genes and pathways specific to epithelial integrity. In the third aim, we will quantify HSV-2 viral shedding over one month in women from the cohort who are HSV-2 seropositive (n=90). In an exploratory analysis, we will also examine the vaginal virome by metagenomic sequencing. The proposed research is significant because it may identify new therapeutic targets for millions of girls and women with FGS. Further, it advances novel studies of parasites and viruses to expand our understanding of interactions between helminths, mucosal immunity, and viral infections.