Natural History of HPV Infection to Neoplasia

描述(申请人提供)：这是我们的“人乳头瘤病毒自然病史；感染至肿瘤”奖助金的竞争性续期。HPV的自然病程很可能受到先天性和获得性粘膜免疫的影响。更具体地说，我们假设Tolllike受体(TLRs)通过分泌促炎、趋化和抗病毒细胞因子在宫颈对HPV的先天免疫中发挥重要作用。上调TLR的表达还将导致树突状细胞和T细胞的激活，进而通过分泌几种细胞因子促进Th1样反应，从而诱导成功的细胞介导免疫(CMI)反应。我们建议：1)在宫颈细胞样本中，检测TRL表达、介导先天免疫的TRL相关细胞因子(干扰素α、肿瘤坏死因子、IL-6和IL-8)与HPV感染清除之间的关系；2)在宫颈细胞样本中，检测TRL表达、诱导和介导获得性免疫的TRL相关细胞因子(干扰素-γ、IL-12、23和27)与HPV清除之间的关系；以及3)检测TLR诱导的Th-1反应、在外周血中检测到的HPV特异性CMI反应和HPV清除之间的关系。A)在1990-1995年初期进入队列并继续受到跟踪的青少年和年轻妇女，以及b)在上一次招募浪潮(2000-2005年)进入队列的青少年和年轻妇女将被要求继续后续行动五年(2005-2010年)。在这项研究开始之时，这些妇女在入院就诊时就已经具备了HPV的特征，每隔4个月对HPVDNA、细胞学、细菌性阴道病、阴道镜检查(宫颈成熟度评估)、沙眼衣原体和淋球菌检测、RT-PCR检测宫颈细胞因子(干扰素-γ、白介素4、10和12)以及HPV16阳性妇女的外周血CMI进行检测。在整个后续行动期间，将继续以同样的时间间隔确定妇女的上述特征。通过使用宫颈细胞的逆转录聚合酶链式反应和Luminex技术使用宫颈粘液的天然和获得性免疫(TLRs，ILS-6，ILS-6，5，13，23和27，肿瘤坏死因子，干扰素)的测量将被添加到与HPV DNA，细胞学和其他宫颈细胞因子描述的相同的4个月间隔检测中。HPV16阳性的妇女将获得额外的CMI血液，使用干扰素-伽马ELISPOT技术检测抗E6和E7反应。我们承认这种设计简化了细胞因子的多效性。然而，我们认为这个模型反映了HPV控制的合理机制，并在我们的队列中进行测试是可行的。从这类研究中获得的信息将对开发疫苗策略和治疗方法以及阐明粘膜上皮中形成的免疫反应至关重要。

DESCRIPTION (provided by applicant): This is a competitive renewal for our "Natural history of HPV; infection to neoplasia" grant. The natural history of HPV is most likely influenced by both innate and adaptive mucosal immunity. More specifically, we hypothesize that Tolllike receptors (TLRs) play an important role in cervical innate immunity to HPV through secretions of proinflammatory, chemotactic and anti-viral cytokines. Up-regulated TLR expression will also result in activation of dendritic cells and T cells that in turn will promote a Thl like response through secretion of several cytokines and consequently, the induction of a successful cell mediated immune (CMI) response. We propose to: 1) examine, in cervical cell samples, the association among TRL expression,TRL-associated cytokines that mediate innate immunity ( IFN alpha, TNF, IL-6 and IL-8) and clearance of incident HPV infection; 2) examine, in cervical cell samples, the association among TRL expression, TRL-associated cytokines that induce and mediate adaptive immunity (IFN-gamma, ILs- 12, 23, and 27) and HPV clearance; and 3) examine the association among TLR induced Th-1 responses measured in cervical cell samples, HPV specific CMI responses detected in peripheral blood (PB) and HPV clearance. Adolescent and young women who were a) entered into the cohort during the initial 1990-1995 period and have continued to be followed and b) entered into the cohort during the last recruitment wave (2000-2005) will be asked to continue follow-up for an additional five years (2005-2010). These women will have been well characterized at the time of the initiation of this study with HPV at their entry visit and 4-month interval sampling for HPV DNA, cytology, bacterial vaginosis, colpophotographs (assessment of cervical maturation), C. trachomatis and N. gonorrohea testing, cervical cell cytokines by RT-PCR (IFN-gamma, ILs 4,10 and 12) and PB CMI for HPV 16 positive women. Women will be continued to be characterized for the above at the same intervals through-out the follow-up. Measures of innate and adaptive immunity (TLRs, ILs-6, 8, 5, 13, 23, and 27, TNF, IFN alpha) by RT PCR using cervical cells and by Luminex technology using cervical mucous will be added to the same 4 month interval testing as HPV DNA, cytology and other cervical cytokines described. Women positive for HPV 16 will get additional blood for CMI using IFN-gamma EliSpot technique for detection of anti-E6 and E7 responses. We acknowledge that this design simplifies the pleiotropic nature of cytokines. However, we feel that this model reflects plausible mechanisms involved in HPV control and is feasible to test in our cohort. Information garnered from this type of study will be critical in developing vaccine strategies and therapies as well as illuminating immune responses developed in the mucosal epithelium.