Reprogramming Host Immune Responses to Cure or Control Persistent HPV Infection

描述（由申请人提供）：持续、慢性人乳头瘤病毒 (HPV) 感染可导致多种疾病，包括复发性呼吸道乳头状瘤病 (RRP)（通常由 HPV 6 或 11 引起）；宫颈上皮内瘤变、宫颈癌、阴茎癌和肛门癌（主要由 HPV 16 或 18 引起）；和口咽癌（HPV16）。这些疾病的免疫研究都无法解释为什么只有一小部分感染这些普遍存在的 HPV 的个体未能遏制/消除其感染。持续活动性感染是HPV诱发恶性肿瘤的主要危险因素。我们对 RRP 进行了广泛的研究，其特点是癌前肿瘤反复生长，需要频繁进行手术，可能因气道完全闭塞或癌症而导致死亡，每年的治疗费用超过 1 亿美元。我们发现 RRP 患者具有 HPV 特异性 TH2 样/Treg 免疫表型，且明显缺乏 TH17 样细胞，并且无法释放促炎细胞因子 IL-36？乳头状瘤强烈表达。 RRP 患者还在气道中持续、强烈地表达环氧合酶-2 (COX-2)，这可能会导致免疫反应出现偏差。我们的初步数据表明，使用塞来考昔（一种 COX-2 抑制剂）治疗可使 RRP 中血清 TH2 样趋化因子正常化，并导致长期、可持续的临床改善；然而，其机制尚不清楚。我们重点关注这种干预策略如何消除/遏制持续性 HPV 感染，并测试我们的新假设，即在 RRP 中重新平衡 HPV 特异性 Treg/TH17 适应性免疫可能会消除/控制持续性 HPV 感染。具体目标将检验以下假设：通过增强 HPV 持续感染组织中的 TH17/Treg 平衡，可以重新编程允许的、无效的抗 HPV 免疫反应，从而导致适应性反应重新极化，转向可以消除活动性疾病的 TH1 样表型。目的是： 1) 确定 IL-36 的释放是否需要 IL-17？来自HPV感染的喉部角质形成细胞，是否有IL-36？可以激活/成熟来自 RRP 患者的朗格汉斯细胞； 2) 确定来自 RRP 患者的 Tregs 是否可以通过连接芳烃 (AHR) 受体而被重新编程为 TH17 样 T 细胞； 3) 确定 TH17 样 T 细胞是否由 IL-36 诱导？激活的 LC 或连接 AHR 后，可以成为 HPV 特异性 TH1 样 T 细胞，支持细胞毒性 TC1 样 T 细胞的生成。这些研究将测试新的概念，即重新调节持续性 HPV 感染患者的病原体特异性免疫反应，模拟其功能，就像大多数具有潜伏 HPV 感染且没有表现出疾病迹象的 HPV 感染者一样。我们正在进行的独特的塞来昔布临床试验可以将体外研究与体内反应进行直接比较，并将为使用这种方法治疗其他持续性 HPV 感染提供依据。这些研究还可以为为什么一些生殖道和口咽部感染高危 HPV 的患者最终发展为恶性肿瘤提供关键且迫切需要的见解。

DESCRIPTION (provided by applicant): Persistent, chronic human papillomavirus (HPV) infection is responsible for multiple diseases including recurrent respiratory papillomatosis (RRP) (usually caused by HPV 6 or 11); cervical intraepithelial neoplasia, cervical, penile and anal cancers (predominantly caused by HPV 16 or 18); and cancers of the oropharynx (HPV16). None of the immune studies of these diseases explain why only a subset of individuals infected with these ubiquitous HPVs fail to contain/eliminate their infection. Persistent active infection i the major risk factor for HPV-induced malignancy. We have extensively studied RRP, characterized by repeated growth of pre-malignant tumors that requires frequent surgeries that can cause mortality due to complete airway occlusion or cancer, and costs >100 million USD to treat/yr. We found that RRP patients have an HPV- specific TH2-like/Treg immunophenotype with a notable absence of TH17-like cells, and a failure to release the pro-inflammatory cytokine IL-36? robustly expressed by papillomas. RRP patients also constitutively, robustly express cyclooxygenase-2 (COX-2) in the airway that can bias immune responsiveness. Our preliminary data show that treatment with celecoxib, a COX-2 inhibitor, normalizes serum TH2-like chemokines in RRP, and causes long-term, sustainable clinical improvement; however, the mechanism is unknown. We focus on how this interventional strategy eliminates/contains persistent HPV infection and we test our novel hypothesis that rebalancing HPV-specific, Treg/TH17 adaptive immunity in RRP may eliminate/control persistent HPV infection. Specific Aims will test the hypothesis that a permissive, ineffective anti-HPV immune response can be reprogrammed through enhancement of the TH17/Treg balance in HPV persistently infected tissues, resulting in re-polarization of the adaptive response toward a TH1-like phenotype that can eliminate active disease. The aims are: 1) Determine whether IL-17 is required for release of IL-36? from HPV-infected laryngeal keratinocytes, and whether IL-36? can activate/mature Langerhans cells from RRP patients; 2) Determine if Tregs from RRP patients can be reprogrammed to become TH17-like T-cells through ligation of their aryl hydrocarbon (AHR) receptor; and 3) Determine if TH17-like T-cells induced by IL-36? activated LCs, or following ligation of their AHR, can become HPV-specific TH1-like T-cells that can support the generation of cytotoxic TC1-like T-cells. These studies will test the novel concept that remodulating pathogen-specific, immune responses of patients with persistent HPV infection simulate to function like most HPV-infected individuals who have latent HPV infection and show no signs of disease. Our unique ongoing clinical trial of celecoxib allows for a direct comparison of in vitro studies with in vivo responses and will provide a rationale to use this approach to treat other persistent HPV infections. These studies could also provide critical and urgently needed insight into why some patients infected with high risk HPVs of the genital tract and oropharynx ultimately develop malignancy.