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

• 批准号: 8636983
• 负责人: Caroline M Mitchell
• 金额: $ 12.71万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8636983
• 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-1; 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; cervicovaginal; 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; pyrosequencing; rRNA Genes; receptor; reproductive; research study; respiratory; response; toll-like receptor 4

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）是一种以阴道厌氧细菌过度生长为特征的阴道综合征，与多种不良临床结果相关，包括盆腔炎、早产和HIV-1感染风险增加。个体之间的细菌种类不同，临床症状、严重后遗症的发生率和细胞因子反应的模式也不同。我们假设临床差异是由个体物种和宿主免疫反应之间的相互作用造成的。我们计划测试以下假设：1)一些细菌物种下调先天宿主反应，通过降低阴道细胞因子和/或抗菌肽（β -防御素、抗菌肽、分泌性白细胞蛋白酶抑制剂[SLPI]、乳铁蛋白）的水平，允许高水平的定植；2)由于这些抗菌肽的基线水平较低，一些个体更容易被定植；3)乳酸菌和bv相关病原体通过TLR-2等模式识别受体激活先天免疫反应的方式不同；4)由于存在毒力因子或破坏宿主免疫反应的机制，某些细菌物种更容易在上生殖道定植。我们将使用qPCR检测16S rRNA基因，对从30名女性中收集的每日阴道拭子进行前瞻性特征分析，并将其与细胞因子和抗菌肽水平的时间反应相关联（目的1）。阴道上皮细胞培养中的反应将用于确定乳酸菌和bv相关微生物是否以不同的方式激活toll样受体，从而产生不同的细胞因子谱，以及TLR-2激活是否与较高水平的抗菌肽相关（目的2）。子宫切除术后收集的子宫内膜样本将采用宽范围16S rRNA PCR和焦磷酸测序进行鉴定，以检测上生殖道定植，并与同一女性的阴道微生物群进行比较，确定微生物群。此外，将测量宫颈细胞因子和防御分子的水平，以评估它们与上尿路定植的关系（目的3）。这项应用的项目负责人Caroline Mitchell医学博士是一名妇产科医生，她的目标是了解女性生殖道如何抵御细菌病原体，最终目的是开发更好的治疗和预防干预措施。她的导师大卫·弗雷德里克斯博士（Dr. David Fredricks）是一位全国知名的研究者，以使用先进的分子技术更好地描述阴道微生物群而闻名，最近又因其卓越的指导而获得认可。华盛顿大学是研究经费最多的公立大学，拥有国际知名的传染病项目和悠久的细菌性阴道炎研究历史。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。

项目成果

Bacterial vaginosis and the cervicovaginal immune response.

• DOI: 10.1111/aji.12264
• 发表时间: 2014-06
• 期刊: American journal of reproductive immunology (New York, N.Y. : 1989)
• 作者: Mitchell C;Marrazzo J
• 通讯作者: Marrazzo J

Colonization of the upper genital tract by vaginal bacterial species in nonpregnant women.

• DOI: 10.1016/j.ajog.2014.11.043
• 发表时间: 2015-05
• 期刊: American journal of obstetrics and gynecology
• 影响因子: 9.8
• 作者: Mitchell CM;Haick A;Nkwopara E;Garcia R;Rendi M;Agnew K;Fredricks DN;Eschenbach D
• 通讯作者: Eschenbach D

Pelvic inflammatory disease: current concepts in pathogenesis, diagnosis and treatment.

• DOI: 10.1016/j.idc.2013.08.004
• 发表时间: 2013-12
• 期刊: Infectious disease clinics of North America
• 影响因子: 4.4
• 作者: Mitchell C;Prabhu M
• 通讯作者: Prabhu M

Hydrogen Peroxide-Producing Lactobacilli Are Associated With Lower Levels of Vaginal Interleukin-1β, Independent of Bacterial Vaginosis.

产生过氧化氢的乳杆菌与较低水平的阴道白细胞介素1β相关，与细菌性阴道病无关。

• DOI: 10.1097/olq.0000000000000298
• 发表时间: 2015-07
• 期刊: Sexually transmitted diseases
• 影响因子: 3.1
• 作者: Mitchell C;Fredricks D;Agnew K;Hitti J
• 通讯作者: Hitti J

High Prevalence of Sexual Dysfunction in a Vulvovaginal Specialty Clinic.

• DOI: 10.1097/lgt.0000000000000085
• 发表时间: 2016-01
• 期刊: Journal of lower genital tract disease
• 影响因子: 3.7
• 作者: Gordon D;Gardella C;Eschenbach D;Mitchell CM
• 通讯作者: Mitchell CM