Female Genital Innate Immune Response to Vaginal Microbiota

女性生殖器对阴道微生物群的先天免疫反应

基本信息

批准号：
8049622
负责人：
Caroline M Mitchell
金额：
$ 12.71万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8049622
关键词：
Advisory Committees Affect Age Anaerobic Bacteria Anti-Inflammatory Agents Anti-inflammatory Bacteria Bacterial Vaginosis Biological Assay Biological Markers Blocking Antibodies Cell Culture System Cell Culture Techniques Cell Line Cells Cervical Clinical Clinical Data Clinical Trials Clostridium Communicable Diseases Communities Complex Cross-Sectional Studies Cytokine Gene Data Data Analyses Defensins Development Down-Regulation Embryo Endometrial Enzyme-Linked Immunosorbent Assay Epithelial Epithelial Cells Female Firefly Luciferases Frequencies Funding Future Gardnerella vaginalis Generations Genital system Goals Gram-Positive Cocci HIV HIV-1 Health High Risk Woman Human Hysterectomy IL8 gene Immune Immune response Immunologic Techniques Incidence Individual Inflammation Inflammatory Inflammatory Response Interleukin-10 Interleukin-17 Interleukin-4 Interleukin-6 Intervention Irrigation Lactobacillus Lactoferrin Left Link Liquid substance Measures Mechanics Mentors Methods Microbe Molecular Morbidity - disease rate Mucous Membrane Mucous body substance Mus NF-kappa B Neutrophil Infiltration Operative Surgical Procedures Oral Outcome Pathway interactions Pattern Pattern recognition receptor Pelvic Inflammatory Disease Peptides Play Predisposition Premature Birth Prevention Preventive Intervention Probiotics Production Receptor Activation Recording of previous events Recruitment Activity Reporter Genes Research Research Personnel Ribosomal RNA Risk Risk Factors Role Sampling Specimen Sterility Surface Swab Symptoms Syndrome TNF gene Techniques Testing Therapeutic Therapeutic Intervention Time Tissues Toll-Like Receptor 1 Toll-Like Receptor 2 Toll-like receptor 6 Toll-like receptors United States Universities Uterus Vagina Vaginal Discharge Virulence Factors Washington Woman antileukoprotease antimicrobial peptide beta-Defensins beta-defensin-2 cathelicidin commensal microbes cytokine enhancing factor gastrointestinal human SLPI protein interleukin-23 kidney cell macrophage microbial microbial community monocyte mucinase neutrophil pathogen pathogenic bacteria prevent programs rRNA Genes receptor reproductive research study respiratory response toll-like receptor 4

项目摘要

DESCRIPTION (provided by applicant): Bacterial vaginosis (BV) is a vaginal syndrome characterized by an overgrowth of anaerobic vaginal bacteria, and is associated with several adverse clinical outcomes, including pelvic inflammatory disease, preterm birth and increased risk of HIV-1 acquisition. The individual bacterial species differ between individuals, as do the clinical symptoms, the incidence of serious sequelae, and patterns of cytokine response. We hypothesize that clinical differences result from the interactions between individual species and host immune responses. We plan to test the following hypotheses: 1) that some bacterial species downregulate the innate host response, allowing high levels of colonization by decreasing levels of vaginal cytokines and/or production of antimicrobial peptides(beta-defensins, cathelicidin, secretory leukocyte protease inhibitor[SLPI], lactoferrin); 2) that some individuals are more vulnerable to colonization due to low baseline levels of those antimicrobial peptides; 3) that Lactobacillus species and BV-associated pathogens activate the innate immune response differently via pattern-recognition receptors like TLR-2; and 4) that some bacterial species are more likely to colonize the upper genital tract due to the presence of virulence factors or mechanisms for subverting the host immune response. We will characterize prospectively collected daily vaginal swabs from 30 women for levels of individual bacterial species using qPCR for the 16S rRNA gene, and correlate these with the temporal response in levels of cytokines and antimicrobial peptides (Aim 1). Responses in vaginal epithelial cell culture will be used to determine whether lactobacilli and BV-associated microbes activate toll-like receptors differently resulting in different cytokine profiles, and whether TLR-2 activation is associated with higher levels of antimicrobial peptides (Aim 2). Endometrial samples collected at hysterectomy will be characterized by broad range 16S rRNA PCR and pyrosequencing to detect upper genital tract colonization and define the microbiota in comparison to the vaginal microbiota in the same women. In addition, levels of cervical cytokines and defense molecules will be measured to assess their relationship with upper tract colonization (Aim 3). The PI for this application, Caroline Mitchell, MD is an OB/Gyn whose goal is to understand how the female genital tract defends against bacterial pathogens, with the ultimate endpoint of developing better therapeutic and preventive interventions. Her mentor, Dr. David Fredricks, is a nationally recognized investigator known for using advanced molecular techniques to better characterize the vaginal microbiota, and has recently been recognized for excellence in mentoring. The University of Washington is the top public university for research funding, with an internationally-renowned Infectious Diseases program and a long history of studies on BV. The scientific advisory committee has expertise in immunologic techniques, clinical trials and data analysis. Narrative: Bacterial vaginosis is a common clinical syndrome that affects up to 30% of women in the United States and is associated with a 2-fold increase of the risk of preterm birth, pelvic inflammatory disease and HIV-1 acquisition. This project will increase our understanding how certain bacterial species change the risk of acquiring bacterial vaginosis and what components of the immune response are most important for protection against bacterial vaginosis. This will allow development of targeted preventive and therapeutic interventions to reduce the adverse clinical outcomes.

描述（由申请人提供）：细菌性阴道病（BV）是一种阴道综合征，其特征是厌氧性阴道细菌过度生长，与几种不良临床结局有关，包括骨盆炎性疾病，早产和增加HIV-1的风险。个体之间的单个细菌物种不同，临床症状，严重后遗症的发生率和细胞因子反应的模式也有所不同。我们假设临床差异是由单个物种与宿主免疫反应之间的相互作用引起的。我们计划检验以下假设：1）一些细菌物种下调了先天的宿主反应，从而通过降低阴道细胞因子水平和/或产生抗菌肽（β-苯二他蛋白酶，凝糖蛋白，athelicidin，separentory Leukocyte蛋白酶蛋白酶蛋白酶抑制剂[slppi]，lactoferrin，lactoferrin，lactoferrin），允许高水平的定殖。 2）由于这些抗菌肽的基线水平较低，有些人更容易受到定殖； 3）乳杆菌物种和与BV相关的病原体通过TLR-2（例如TLR-2）以不同的方式激活先天免疫反应。 4）由于存在毒力因子或机制来颠覆宿主免疫反应，因此某些细菌物种更有可能在上生殖道中定居。我们将使用QPCR进行16S rRNA基因的三名女性的前瞻性收集的每日阴道拭子，以介绍单个细菌种类的水平，并将其与细胞因子和抗菌肽水平的时间反应相关联（AIM 1）。阴道上皮细胞培养的反应将用于确定乳酸杆菌和与BV相关的微生物激活类Toll样受体是否不同，从而导致不同的细胞因子谱，以及TLR-2激活是否与较高水平的抗菌肽相关（AIM 2）。在子宫切除术中收集的子宫内膜样品的特征是宽范围16S rRNA PCR和pyrosequencing，以检测上生殖道的定殖并定义与同一女性的阴道微生物群相比定义菌群。此外，将测量宫颈细胞因子和防御分子的水平来评估它们与上层定殖的关系（AIM 3）。该应用的PI，医学博士Caroline Mitchell是妇产科，其目标是了解女性生殖道如何防御细菌病原体，并具有开发更好的治疗和预防性干预措施的最终终点。她的导师戴维·弗雷德里克斯（David Fredricks）博士是一位全国认可的调查员，以使用先进的分子技术来更好地描述阴道微生物群，并因在指导方面的卓越表现而受到认可。华盛顿大学是研究资助的顶级公立大学，拥有国际知名的传染病计划，并且在BV方面有悠久的研究历史。科学咨询委员会在免疫技术，临床试验和数据分析方面具有专业知识。叙事：细菌性阴道病是一种常见的临床综合征，影响美国30％的妇女，并且与早产，骨盆炎性疾病和HIV-1获取的风险增加了2倍。该项目将加深我们的理解，某些细菌种类如何改变获得细菌性阴道病的风险，以及免疫反应的哪些组成部分对于防止细菌性阴道病的保护最为重要。这将允许开发有针对性的预防和治疗性干预措施，以减少不良临床结果。