Development of immune assays for HPV-32

HPV-32 免疫检测的开发

• 批准号: 6880090
• 负责人: MICHAEL E HAGENSEE
• 金额: $ 7.1万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6880090
• 关键词: HIV infections; T lymphocyte; capsid; clinical research; communicable disease diagnosis; diagnostic tests; enzyme linked immunosorbent assay; flow cytometry; genotype; high throughput technology; human papillomavirus; human subject; humoral immunity; immune response; molecular cloning; opportunistic infections; oral health; patient oriented research; serology /serodiagnosis; virus antigen; virus diseases; virus genetics; warts

DESCRIPTION (provided by applicant): Opportunistic infections of the oral cavity afflict 50% of all HIV infected patients, and include oropharyngeal candidiasis (OPC), oral hairy leukoplakia (OHL) and oral warts caused by the mucosatropic human papillomavirus (HPV). The aggressive treatment of HIV with highly-active antiretroviral therapy (HAART) has significantly improved the HIV patient's health and prognosis by lowering systemic HIV viral loads and restoring immune function primarily through increases in CD4+ T lymphocytes. This has resulted in a substantial decrease in the incidence of HIV-associated opportunistic oral diseases, including OPC and (OHL). In stark contrast, the incidence of oral papillomas (warts) has reportedly increased since the widespread administration of HAART. Preliminary analysis has identified HPV-32 as the predominant genotype found in oral warts in the New Orleans HIV+ cohort. It is felt that asymptomatic oral HPV infection occurs frequently but oral warts occur infrequently, presumably due to immunological control of the virus. The critical aspects of the immune response that prevent the progression from asymptomatic HPV infection to HPV disease are unknown, but previous studies have focused on HPV genotype-specific response against the major capsid protein, LI. The increased rate of HPV-related oral pathology seen in HIV+ patient's highlights the need for a more thorough understanding of the immune response to oral HPV infections. Furthermore, the accessibility of the oral cavity affords a unique opportunity to conduct rigorous analysis of HPV infection, immunity and pathogenesis. To initiate these studies, the development of the immunological assays specific for oral HPV genotypes such as HPV-32 are required. We hypothesize that HPV-32 specific humoral and cellular immunoassays can be developed and utilized to screen populations for HPV-32 specific immune responses. We propose to develop an enzyme-linked immunosorbent assay (ELISA) for detection of HPV-32-specific antibodies in serum, and lymphoproliferative and flowcytometric assays for the detection of HPV-32-specific T cell responses. These assays will be tested for optimal assay conditions, specificity, and reproducibility. Ultimately these assays will be used to investigate the role of immunity in the acquisition and subsequent clearing or progression of oral HPV infections, particularly in the highly susceptible HIV+ patient. The goal of this project is to develop the tools necessary to gain a better understanding of the humoral and cellular immune response to oral HPV infection, particularly in the HIV+ individual. The understanding of immune response to oral HPV infection may help to predict HPV disease progression as well as elucidate therapeutic and preventive strategies for HPV-related disease.

描述（由申请人提供）：口腔的机会感染遭受了所有艾滋病毒感染患者的50％，包括口咽念珠菌病（OPC），口腔毛状白血病（OHL）和由粘膜胶质性人乳头虫（HPV）引起的口腔疣。高度活跃抗逆转录病毒疗法（HAART）对HIV的积极治疗通过降低全身HIV病毒载荷和主要通过CD4+ T淋巴细胞的增加来显着改善HIV患者的健康和预后。这导致包括OPC和（OHL）在内的HIV相关机会性口腔疾病的发生率大大降低。相比之下，据报道，自哈特（Haart）广泛管理以来，口服乳头瘤（疣）的发病率有所增加。初步分析已将HPV-32鉴定为在新奥尔良HIV+队列中口腔疣中发现的主要基因型。人们认为无症状的口服HPV感染经常发生，但口腔疣很少发生，大概是由于对病毒的免疫控制。免疫反应的关键方面是防止无症状HPV感染对HPV疾病的进展的关键方面，但以前的研究集中在HPV基因型特异性反应上，针对主要的capsid蛋白Li。 HIV+患者中与HPV相关的口腔病理学率的提高突显了需要对口服HPV感染的免疫反应进行更彻底的了解。此外，口腔的可及性为对HPV感染，免疫和发病机理进行严格分析提供了独特的机会。为了启动这些研究，需要开发针对口服HPV基因型（例如HPV-32）的特定的免疫学测定。我们假设可以开发并利用HPV-32特定的体液和细胞免疫测定，用于筛选HPV-32特异性免疫反应的种群。我们建议开发一种酶联免疫吸附测定法（ELISA），以检测血清中HPV-32特异性抗体，以及用于检测HPV-32特异性T细胞反应的淋巴增生性和流环测定法。这些测定将测试最佳测定条件，特异性和可重复性。最终，这些测定法将用于研究免疫在获得和随后的口服HPV感染的清除或进展中的作用，特别是在高度易感的HIV+患者中。 该项目的目的是开发必要的工具，以更好地了解对口服HPV感染的体液和细胞免疫反应，尤其是在HIV+个体中。对口服HPV感染的免疫反应的理解可能有助于预测HPV疾病的进展，并阐明与HPV相关疾病的治疗和预防策略。