Mechanisms of mucosal immune evasion in high grade cervical dysplasia

描述（由申请人提供）：这项工作的目标是研究归巢机制，提供免疫监视和生殖器粘膜的稳态。这些研究是基于一系列由NIH资助的临床方案，测试治疗性疫苗用于治疗高度宫颈不典型增生（CIN 2/3）的妇女，这种病变是浸润性宫颈癌的直接前兆。CIN 2/3是一种对HPV特异性T细胞反应敏感的疾病。HPV 16 E6和E7的抗原靶标是非“自身”的，并且在功能上是疾病起始和持续所需的。CIN 2/3病变相对容易，其中一些确实会消退。这一提议背后的具体假设是，CIN 2/3病变减轻了局部免疫效应细胞消除疾病的能力，并且可以操纵病变微环境以实现基于免疫的治疗成功。这项工作将分析来自一系列免疫启动试验的临床标本，使用我们机构的同事开发的治疗试剂，由NCI RAID（pNGVL 4a-sig/E7 /HSP 70）生产，并作为CelticPharma（TA-HPV）的礼物。这项工作的长期目标是确定导致HPV特异性适应性反应未能根除CIN 2/3的免疫逃逸机制，确定预测免疫治疗反应的生物标志物，并开发联合干预措施以解偶联免疫逃逸机制。具体而言，我们将：SA1：确定宫颈T细胞上的归巢受体谱和正常和CIN 2/3粘膜中相应配体的表达。宫颈粘膜归巢机制的表征将允许监测可能运输到生殖道的免疫应答，可能提示有效的免疫途径，例如，口腔或鼻腔或生殖器的致敏，以及在我们可以确定特异性的情况下，也可以使我们估计可检测的全身免疫应答与病变部位的测量相关的程度。SA2：确定持续性CIN 2/3的免疫学特征：我们将确定正常宫颈粘膜和CIN 2/3中常驻免疫细胞群的表型、功能潜力、分布和共定位，确定与粘膜上皮和间质区室中存在或不存在CD 8+浸润相关的趋化因子/趋化因子受体谱，并确定T0时对疫苗接种有反应的受试者和对咪喹莫特有反应的受试者与无反应的受试者的趋化因子谱。 公共卫生相关性：高度宫颈发育不良中的粘膜免疫逃避机制HPV相关疾病中生殖器免疫应答的归巢机制的鉴定可能提示最佳的疫苗接种途径，为可能运输到生殖器粘膜的免疫应答的监测提供信息，并确定宫颈中的免疫应答可以检测到的程度或与外周血中的测量相关。从恶性肿瘤的T细胞疗法开发的观点来看，由于发育不良的宫颈病变相对容易获得，并且许多人实体瘤常见的病变介导的免疫抑制机制可能区分应答者和非应答者，因此这些研究提供了测试免疫治疗组合的原理证明的机会。

DESCRIPTION (provided by applicant): The goal of this work is to study mechanisms of homing that provide immune surveillance and homeostasis in the genital mucosa. The studies are based on a series of investigator-initiated clinical protocols which are funded by the NIH, testing therapeutic vaccines for the treatment of women with high grade cervical dysplasia (CIN2/3), the lesion which is the immediate precursor to invasive cervical cancer. CIN2/3 is a disease which should be susceptible to an HPV-specific T cell response. The antigenic targets, HPV16 E6 and E7, are non- 'self', and functionally required for disease initiation and persistence. CIN2/3 lesions are relatively accessible, and some of them do regress. The specific hypothesis behind this proposal is that CIN2/3 lesions mitigate the ability of localized immune effector cells to eliminate disease, and that the lesion microenvironment can be manipulated to enable immune-based therapeutic success. This work will analyze clinical specimens from a series of investigator-initiated trials, using therapeutic reagents developed by colleagues at our institution, and manufactured by NCI RAID (pNGVL4a-sig/E7 /HSP70), and as a kind gift from CelticPharma (TA-HPV). The long term goals of this work are to identify mechanisms of immune escape which contribute to the failure of HPV-specific adaptive responses to eradicate CIN2/3, to identify biomarkers predictive of response to immune-based therapies, and to develop combinatorial interventions to uncouple immune escape mechanisms. Specifically, we will: SA1: Determine the homing receptor profile on cervical T cells and expression of corresponding ligands in normal and CIN2/3 mucosa. The characterization of homing mechanisms for cervical mucosa will allow monitoring of immune responses that are likely to traffic to the genital tract, may suggest efficient routes of immunization, e.g., oral or nasal or genital priming, and in cases in which we can determine specificity, may also allow us to estimate the extent to which detectable systemic immune responses correlate with measures in the lesion site. SA2: Determine the immunologic signature of persistent CIN2/3: We will determine the phenotype, functional potential, distribution and co localization of resident immune cell populations in normal cervical mucosa and in CIN2/3, determine the chemokine/chemokine receptor profile associated with presence or absence of CD8+ infiltration in mucosal epithelial and stromal compartments, and determine the chemokine profile at T0 of subjects who respond to vaccination, and of subjects who respond to imiquimod, compared to those who do not. PUBLIC HEALTH RELEVANCE: Mechanisms of mucosal immune evasion in high grade cervical dysplasia The identification of homing mechanisms for genital immune responses in HPV-associated disease is likely to suggest optimal routes of vaccination, to inform monitoring of immune responses likely to traffic to the genital mucosa, and to determine the extent to which immune responses in the cervix can be detected or correlated with measurements in the peripheral blood. From the standpoint of development of T cell therapies for malignancies, because dysplastic cervical lesions are relatively accessible, and mechanisms of lesion- mediated immune suppression common to many human solid tumors are likely to distinguish responders from non-responders, these studies present an opportunity to test proof-of-principle of immune therapeutic combinations.