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在生殖器粘膜中提供免疫监视和动态平衡的归巢机制。这些研究基于一系列由NIH资助的研究人员发起的临床方案，测试用于治疗高度宫颈异型增生(CIN2/3)的女性的治疗性疫苗，CIN2/3是侵袭性宫颈癌的直接先兆。CIN2/3是一种疾病，应该容易受到HPV特异性T细胞反应的影响。抗原靶点HPV16E6和E7是非“自身”的，在功能上是疾病启动和持续所必需的。CIN2/3的病变相对容易，有些确实会消退。这一建议背后的特定假设是，CIN2/3病变降低了局部免疫效应细胞消除疾病的能力，并且病变微环境可以被操纵，以实现基于免疫的治疗成功。这项工作将分析一系列由研究人员发起的试验的临床标本，使用由我们机构的同事开发的治疗试剂，并由NCI RAID(pNGVL4a-sig/E7/HSP70)制造，并作为CelticPharma(TA-HPV)的礼物。这项工作的长期目标是确定导致HPV特异性适应性反应失败的免疫逃逸机制，以根除CIN2/3，识别预测免疫治疗反应的生物标志物，并开发组合干预措施来解偶联免疫逃逸机制。具体地说，我们将：SA1：确定宫颈T细胞上的归巢受体特征以及相应的配体在正常和CIN2/3粘膜中的表达。宫颈黏膜归巢机制的特征将使我们能够监测可能流向生殖道的免疫反应，可能建议有效的免疫途径，例如，口服、鼻腔或生殖器启动，在我们可以确定特异性的情况下，还可能允许我们估计可检测到的全身免疫反应与病变部位的措施相关联的程度。SA2：确定持续性CIN2/3的免疫学特征：我们将确定正常宫颈黏膜和CIN2/3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间CD8+是否渗入相关的趋化因子/趋化因子受体谱，并确定接种疫苗有反应者和咪喹莫特无反应者T0处的趋化因子谱。 公共卫生相关性：高度宫颈不典型增生的粘膜免疫逃逸机制HPV相关疾病生殖器免疫反应归巢机制的确定可能会建议最佳接种途径，为监测可能进入生殖器粘膜的免疫反应提供信息，并确定宫颈免疫反应可被检测到的程度或与外周血中的测量相关联。从T细胞治疗恶性肿瘤的发展角度来看，由于宫颈发育不良病变相对容易获得，以及许多人类实体肿瘤常见的病变介导的免疫抑制机制可能区分有效和无效，这些研究提供了测试免疫治疗组合的原则证明的机会。

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.