Mucosal immune response of the anus in women to HPV, intercourse, smoking and OCs

描述(由申请人提供)：本申请涉及临床研究的挑战领域(04)；主题：A1-101：开发新方法并解决粘膜免疫学中的关键问题。肛门粘膜免疫学的研究(与所有粘膜上皮一样)具有极大的挑战性，原因有很多：1)肛门具有复杂的微生物环境，这要求对共生生物有一定程度的免疫耐受；2)经常暴露于同时存在的病原体以及肛交引起的创伤；3)缺乏适合于队列研究的技术来研究免疫参数和4)访问 组织和液体的纵向队列通常是困难的。长期以来，肛门粘膜一直被认为容易受到包括艾滋病毒和人乳头瘤病毒(HPV)在内的性传播病原体的感染。当然，更好地了解这些反应将对开发用于性传播感染(STI)预防和治疗干预的局部杀微生物剂至关重要。特别是，需要对HPV等病毒病原体进行治疗干预，HPV是最常见的性传播感染，导致80%-100%的肛门癌。尽管HPV感染肛门很常见，但即使在女性中，大多数感染通常是明确的。不幸的是，对于那些HPV持续存在的人来说，肛门癌的风险仍在继续--这是所有肛门癌发展的关键。相当有力的证据表明，免疫机制影响HPV的清除。了解导致HPV持续存在的免疫学机制对于开发HPV诱导的癌前病变的治疗方法至关重要。此外，众所周知，HPV本身也会增加艾滋病毒的风险。因此，研究人乳头瘤病毒也可能有助于了解性传播感染导致艾滋病毒传播的机制。 在过去的15年里，我们试图通过将免疫分析纳入我们正在进行的纵向队列中：始于1990年的青少年HPV的自然病史(R37 CA51323)，来解决宫颈和阴道的许多免疫学问题。该队列每隔4个月接受一次宫颈共生体和致病微生物的密切特征描述，以及行为访谈。多年来，我们采用了几种适合于队列研究的先天免疫和获得性免疫检测方法，包括检测细胞因子和趋化因子，以及测量宫颈样本中Toll样受体(TLRmRNA)的表达。在这个建立良好的队列中，我们已经存储了自1993年以来每隔4个月收集的超过15,000份肛门样本。我们有一个史无前例的独特机会，可以在一个特征明确的队列中检查一段时间内的粘膜免疫反应。这项研究的目的是：1)在肛门标本中检测在HPV感染时增强的细胞因子；2)在肛门检测 研究对象包括样本、介导先天免疫的细胞因子(干扰素-a、肿瘤坏死因子、白介素6和白介素8)和获得性免疫(干扰素伽马和白介素12)之间的关系，以及HPV感染事件的清除。还将检查可能调节细胞因子和趋化因子谱的辅助因素或行为，包括频繁的肛交、吸烟和口服避孕药的使用；3)比较HPV感染和清除后肛门和宫颈之间的细胞因子和趋化因子谱，以及4)比较HPV感染妇女与肛门细胞学正常和异常的妇女之间的细胞因子/趋化因子谱。我们有超过15,000份储存的肛门样本可用于HPV DNA测试和AIMS 1-3的细胞因子/趋化因子分析。所有储存的肛门样本都将进行HPV DNA检测。细胞因子和趋化因子将从发生HPV感染的探访中获得的样本中进行分析。对于目标3，已经通过父母研究获得了用于HPV DNA和细胞因子/趋化因子分析的妇女宫颈样本以供比较。我们还提出了一个子目标，在这个子目标中，我们将检查感染肛门HPV的女性肛门样本中TLR的表达。由于目前的肛门样本不适合测量TLRs，我们现在已经开始收集 这些样本是前瞻性的。我们预计会有300个样本可供化验。总而言之，我们有独特的机会使用储存的样本在具有良好特征的队列中检查肛门粘膜免疫反应，这将使我们能够严格遵守我们的时间表。这些数据将使我们更好地了解影响粘膜免疫图谱的环境因素。 公共卫生相关性：该应用程序计划检查对人类乳头瘤病毒、烟草使用、肛交和激素避孕药的肛门粘膜免疫反应。

DESCRIPTION (provided by applicant): This application addresses the Challenge area of Clinical research (04); Topic: A1-101: Develop novel methods and address key questions in mucosal immunology. The study of mucosal immunology of the anus (as with all mucosal epithelium) has been extremely challenging for numerous reasons: 1) the anus has a complex microbiologic environment which requires a level of immune tolerance to commensal organisms, 2) there are often frequent exposures to simultaneous pathogens as well as trauma induced by anal intercourse, 3) there has been a lack of techniques suitable to cohort studies to study immune parameters and 4) accessing tissue and fluids in longitudinal cohorts is often difficult. Anal mucosa has long been known to be vulnerable to sexually transmitted pathogens including HIV and human papillomavirus (HPV). Certainly, a greater understanding of these responses will be critical in the development of topical microbicides for sexually transmitted infection (STI) prevention as well as therapeutic interventions. In particular, therapeutic interventions are needed for viral pathogens such as HPV which is the most common STI and is responsible for 80-100% of anal cancers. Although HPV infections of the anus are common, even among women, most infection usually clear. Unfortunately, the risk of anal cancer continues for those with HPV persistence--the key in the development of all anogenital cancers. Fairly strong evidence suggests that immune mechanisms influence HPV clearance. Understanding the immunologic mechanisms that result in HPV persistence is critical in the development of therapeutic treatment for HPV-induced pre-cancers. In addition, HPV itself has been known to increase risks for HIV. Hence studying HPV may also give insight into mechanisms into which STIs contribute to HIV transmission. Over the past 15 years, we have attempted to address many of the immunologic questions in the cervix and vagina by incorporating immune assays in our ongoing longitudinal cohort: the natural history of HPV in teens (R37 CA51323) which began in 1990. This cohort undergoes close characterization for commensal and pathogenic organisms of the cervix, as well as behavioral interviews, at 4-month intervals. Over the years, we have adapted several assays focusing on innate and adaptive immunity that are suitable for study in the cohort including detection of cytokines and chemokines as well as measuring mRNA expression of Toll- like receptors (TLR) from cervical samples. In this well-established cohort, we have stored over 15,000 anal samples which have been collected since 1993 at 4 month intervals. We have the unprecedented and unique opportunity to examine mucosal immune responses over time in a well-characterized cohort. Aims of this study are to: 1) examine, in anal samples, the cytokines which are enhanced at time of HPV acquisition; 2) examine in anal samples, the association between cytokines that mediate innate immunity (IFN-a, TNF, IL-6 and IL-8) and adaptive immunity (IFN gamma and IL 12) and clearance of incident HPV infection. Co-factors or behaviors that may also mediate cytokine and chemokine profiles will be examined including frequent anal intercourse, smoking cigarettes and oral contraceptive use and 3) compare cytokines and chemokine profiles between the anus and the cervix in response to HPV acquisition and clearance, and 4) to compare the cytokine/chemokine profiles between HPV infected women with normal and abnormal anal cytology. We have over 15,000 stored anal samples available for HPV DNA testing and cytokine/chemokine analysis for aims 1-3. All stored anal samples will be tested for HPV DNA. Cytokine and chemokine analysis will be performed from samples obtained from visits with an incident HPV infection. For aim 3, samples from the cervix of women for HPV DNA and cytokine/chemokine analysis are already available through the parent study for comparison. We also propose a subaim in which we will examine TLR expression from anal samples in women who acquire anal HPV. Since the current anal samples are not suitable for measuring TLRs, we have now started collecting these samples prospectively. We expect that we will have 300 samples available for examination for this aim. In summary, we have the unique opportunity to examine anal mucosal immune responses in a well characterized cohort using stored samples which will allow us to keep to our strict timeline. This data will give us a greater understanding of environmental factors that affect mucosal immune profiles. PUBLIC HEALTH RELEVANCE: This application plans to examine anal mucosal immune responses to human papillomavirus, tobacco use, anal intercourse and hormonal contraceptives.