Human Neutrophil Response to Trichomonas vaginalis

人类中性粒细胞对阴道毛滴虫的反应

• 批准号: 9191266
• 负责人: Frances Mercer
• 金额: $ 5.8万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9191266
• 关键词: Acute; Affect; Biological Assay; Bite; Blood; Cell Line; Cells; Cessation of life; Chemicals; Chronic; Clinical; Clinical Research; Coculture Techniques; Cues; Cytolysis; DNA; Data; Drug resistance; Effector Cell; Epithelial; Epithelial Cells; Exposure to; Flow Cytometry; HIV; HIV Infections; Heating; High Prevalence; Human; IL8 gene; Image; Immobilization; Immune; Immune response; Immune system; Immunity; Immunotherapy; In Vitro; Incidence; Infection; Inflammation; Inflammatory; Inflammatory Response; Intervention; Knowledge; Laboratories; Lead; Life; Link; Low Birth Weight Infant; Lytic; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of prostate; Measures; Mediating; Membrane; Monitor; Mus; Neoplasms; Nuclear; Outcome; Parasites; Pathogenesis; Pathology; Patients; Pelvic Inflammatory Disease; Predisposition; Process; Risk; Role; Sexually Transmitted Diseases; Signal Transduction; Site; System; Testing; Therapeutic Intervention; Tissues; Translating; Trichomonas Infections; Trichomonas vaginalis; United States; Vagina; Work; acute symptom; arm; base; cancer complication; cell killing; cell type; cytokine; design; extracellular; fighting; immune clearance; immunogenic; inhibitor/antagonist; killings; live cell imaging; microscopic imaging; mouse model; neutrophil; pathogen; programs; reproductive; resistant strain; response

PROJECT SUMMARY / ABSTRACT Trichomonas vaginalis (Tv) is a protozoan pathogen that causes the most common non-viral sexually transmitted infection in the United States and worldwide. Studies on Tv pathogenesis and immunity have been limited. However, the rise in drug resistant strains of Tv, and an emerging appreciation of the link between asymptomatic Tv infections with inflammation-driven pathologies demands better understanding of this prevalent human infection. Notably, Tv infection has been linked to reproductive complications, increased susceptibility to HIV, increased incidence of cervical cancer, and increased aggressiveness of prostate cancer. Clinical observation, previous work by others, and preliminary studies in our laboratory point to neutrophils (PMN) as the key player in anti-Tv immunity. PMN are extremely destructive cells, propagating a cascade of tissue damage and inflammation, which has been associated with inflammatory pathologies, initiation of new cancers, and exacerbation of existing neoplasms. If an infection is not cleared efficiently, the cycle of inflammation could persist chronically. While the importance of PMN in Tv infection is recognized, how they work to fight Tv infection is not characterized. Therefore, we aim to (i) determine how PMN kill Tv, and (ii) determine the role of PMN in establishing inflammation during Tv infection. Our preliminary data suggest that PMN kill Tv using trogocytosis (trogo= nibble), a process by which effector cells take small “bites” from target cells. We will isolate PMN from human blood and co-culture with Tv. We will then use flow cytometry- based cytolysis assays, live cell imaging microscopy and imaging flow cytometry to determine whether PMN trogocytosis leads to death of Tv. We will also determine whether PMN use neutrophil extracellular traps NETosis to kill Tv, a process by which PMN release nuclear contents externally to ensnare, immobilize and kill pathogens. To assess NETosis, we will use extracellular DNA quantification, imaging, and flow-cytometry based cytolysis assays. In addition, to determine the consequences of PMN-Tv interaction on inflammation, we will probe supernatants of PMN-Tv co-cultures to determine the cytokine program that Tv elicits. We will use live Tv versus heat-inactivated Tv to determine the contribution of Tv-mediated tissue destruction on inflammation, and we will also perform Tv-PMN co-cultures in the presence of vaginal epithelial cells to determine how destruction of the epithelial layer by Tv and PMN contributes to inflammation. The knowledge gained from this study on how acute Tv infection is cleared by PMN and what inflammatory signals are established, will help to suggest potential strategies to monitor inflammatory pathologies, cervical cancers and prostate cancers associated with Tv infection, and inform the design of potential immunotherapy interventions.

项目总结/摘要 阴道毛滴虫（Tv）是一种原生动物病原体，可引起最常见的非病毒性性 在美国和世界范围内传播感染。关于Tv的致病机理和免疫学的研究已经取得了很大进展， 有限公司然而，随着耐药菌株的增加，以及人们对耐药菌株之间的联系的认识不断加深， 无症状的Tv感染与炎症驱动的病理需要更好地了解这一点 流行的人类感染。值得注意的是，Tv感染与生殖并发症有关， 艾滋病毒的易感性，宫颈癌的发病率增加，前列腺癌的侵袭性增加。 临床观察、他人先前的工作以及我们实验室的初步研究都指向中性粒细胞 (PMN)是抗电视免疫的关键人物中性粒细胞是极具破坏性的细胞， 组织损伤和炎症，这与炎症病理学有关，引发新的 癌症和现有肿瘤的恶化。如果感染没有被有效清除， 炎症可能会长期存在。虽然中性粒细胞在Tv感染中的重要性已被认识，但它们如何在Tv感染中发挥作用？ 对抗Tv感染的工作没有特点。因此，我们的目标是（i）确定PMN如何杀死Tv，以及（ii） 确定PMN在Tv感染期间建立炎症中的作用。我们的初步数据显示， PMN通过胞啃作用（trogo= nibble）杀死Tv，这是一个效应细胞从Tv中“咬”出一小口的过程。 靶细胞我们将从人血液中分离PMN并与Tv共培养。我们会用流式细胞术- 基于细胞溶解试验，活细胞成像显微镜和成像流式细胞术，以确定是否PMN 胞啃作用导致Tv死亡。我们还将确定中性粒细胞是否使用中性粒细胞胞外陷阱 NETosis杀死Tv，这是PMN向外释放核内容物以诱捕、捕获和杀死Tv的过程 病原体为了评估NETosis，我们将使用细胞外DNA定量，成像和流式细胞术 基于细胞溶解测定。此外，为了确定PMN-Tv相互作用对炎症的影响， 我们将探测PMN-Tv共培养物的上清液，以确定Tv诱导的细胞因子程序。我们将 使用活Tv与热灭活Tv来确定Tv介导的组织破坏对 炎症，我们还将在阴道上皮细胞存在下进行Tv-PMN共培养， 确定Tv和PMN对上皮层的破坏如何导致炎症。知识 从这项研究中获得了关于急性Tv感染如何被PMN清除以及炎症信号是什么的信息。 建立，将有助于提出潜在的战略，以监测炎症病理，宫颈癌， 与Tv感染相关的前列腺癌，并为潜在的免疫治疗干预措施的设计提供信息。