Interactions between trichomonasviruses, their parasite host, and the human superhost

滴虫病毒、其寄生虫宿主和人类超级宿主之间的相互作用

• 批准号: 10671988
• 负责人: carrie ann Hetzel
• 金额: $ 4.19万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671988
• 关键词: Adherence; Biological Models; Candidate Disease Gene; Cells; Cervical; Clinical; Coculture Techniques; Daughter; Disease Progression; Double Stranded RNA Virus; Double-Stranded RNA; Epithelial Cells; Exonuclease; Gene Expression; Genes; Genetic; High-Throughput RNA Sequencing; Human; IRF3 gene; Infection; Inflammation; Inflammatory; Inflammatory Response; Interferons; Life Cycle Stages; Longitudinal Studies; Malignant neoplasm of prostate; Measures; Mediating; Morphology; Mothers; NF-kappa B; Parasites; Parents; Pathway interactions; Pelvis; Predisposition; Premature Birth; Proliferating; Proteins; RNA; RNA Viruses; RNA-Binding Proteins; Reporting; Role; Severity of illness; Sexually Transmitted Diseases; TLR3 gene; Testing; Tissue-Specific Gene Expression; Trichomonas Infections; Trichomonas vaginalis; Vagina; Viral; Virion; Virulence; Virus; Virus Diseases; Virus Replication; Work; differential expression; experimental study; extracellular; fitness; helicase; human disease; human pathogen; nucleoside analog; response; transcriptome sequencing; transmission process; urogenital tract; virus host interaction

PROJECT SUMMARY Trichomonas vaginalis viruses (TVVs) are double stranded RNA viruses that persist in the obligate human parasite Trichomonas vaginalis (Tvag). Tvag is the causative agent of the human disease trichomoniasis, the most common nonviral sexually transmitted infection worldwide. Infection with TVV-containing parasites is thought to increase the severity of this disease and its complications by increasing the pro-inflammatory response by the human superhost. There are five known species in genus Trichomonasvirus (TVV1, TVV2, TVV3, TVV4, and TVV5), but most experiments to date have been performed using purified TVV1 virions or TVV1-containing isolates, due in part to the greater availability of Tvag isolates singly infected with TVV1. TVVs do not have an extracellular lifecycle and instead are transmitted vertically as the parasite divides. Further, multiple TVV species can coinfect a singular trichomonad. Thus, studying the effect of TVVs on the parasite and the human superhost has been difficult, as uninfected trichomonads cannot be infected to create isogenic TVV+ vs. TVV- strains. Using the nucleoside analog 2’-C-methylcytidine (2CMC), which has been recently reported to clear trichomonads of TVV-infection, we have generated isogenic pairs of cured and uncured strains generated from the same parent isolate. With these isogenic pairs, we propose to test our hypothesis that T. vaginalis maintains a persistent viral infection, which confers a fitness advantage to the protozoan and increases virulence to the human superhost. Thus far, we have performed high throughput RNA-sequencing with one such isogenic pair to measure differential gene expression. In doing so, we have identified several candidate genes that may be involved in limiting viral replication to tolerable levels in order to maintain long-term viral persistence. These differentially expressed genes have largely not been characterized, but some encode factors with homology to known RNA exonucleases and helicases, which we hypothesize might be involved in limiting viral replication. In Aim 1 of this project, we propose to further validate these results in additional isogenic pairs in order to define factors Tvag uses to maintain a stable trichomonasvirus infection. In Aim 2, we propose to determine impacts of TVV infection on Tvag proliferation, morphology, and adherence to human cells. We hypothesize that TVVs will increase parasite survival and attachment, providing an evolutionary explanation for long-term TVV persistence. Finally, in Aim 3, we propose to compare responses of human cells to infection with TVV+ and TVV- trichomonads across TVV species. We anticipate that TVV+ Tvag strains, regardless of virus species, will elicit stronger inflammatory responses than TVV- strains. Through this work, we hope to uncover valuable information about an important human pathogen and an intriguing model system to study virus–host interactions.

项目摘要 阴道病毒（TVVS）是双链的RNA病毒，持续存在于人类 寄生虫阴道（TVAG）。 TVAG是人类疾病毛诺病的病因， 全世界最常见的非病毒性传播感染。感染含TVV的寄生虫的是 被认为通过增加促炎性来增加该疾病的严重程度及其并发症 人类超级主机的反应。 Trichomonasvirus属中有五种已知物种（TVV1，TVV2， TVV3，TVV4和TVV5），但是迄今为止，大多数实验都是使用纯化的TVV1病毒或 含TVV1的分离株，部分原因是TVV1单独感染的TVAG隔离株的可用性更大。 TVVS 没有细胞外生命周期，而是在寄生虫划分时垂直传播。此外， 多种TVV物种可以共同感染一个单一的毛刺。这是研究TVV对寄生虫的影响以及 人类的超主机很困难，因为未感染的毛毛纳德不能感染来创建等源性TVV+ 与TVV-Strewns。使用核苷类似物2'-C-甲基胞丁氨酸（2CMC），最近已报道 明确的TVV感染的毛trichomonads，我们产生了固化和未固化的菌株的等异生对 来自同一父孤立。通过这些等源性对，我们提出了我们的假设，即阴道t. 保持持续的病毒感染，该病毒感染了原生动动的健身优势并增加 对人类超级主机的毒力。那遥远，我们已经用一个 这样的等源性对测量差异基因表达。这样，我们已经确定了几个候选人 可能涉及将病毒复制限制为可耐受水平以维持长期病毒的基因 持久性。这些不同表达的基因在很大程度上没有被表征，但是某些编码因子 与已知的RNA外丝酶和解旋酶同源，我们认为这可能参与限制 病毒复制。在该项目的AIM 1中，我们建议在其他同源对中进一步验证这些结果 为了定义TVAG用途的因素，以维持稳定的Trichomonasvirus感染。在AIM 2中，我们建议 确定TVV感染对TVAG增殖，形态和对人类细胞的依从性的影响。我们 假设TVV将增加寄生虫的生存和依恋，从而为 长期TVV持久性。最后，在AIM 3中，我们建议将人类细胞的反应与感染的反应与 TVV+和TVV- TVV-TVV物种。我们预计TVV+ TVAG菌株，无论病毒如何 物种将引起比TVV-Strewns更强烈的炎症反应。通过这项工作，我们希望发现 关于重要人类病原体和研究病毒 - 主机的有趣模型系统的有价值的信息 互动。