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）：50%的HIV感染患者患有口腔念珠菌感染，包括口咽念珠菌病（OPC）、口腔毛状白斑（OHL）和由粘膜萎缩性人乳头瘤病毒（HPV）引起的口腔疣。用高效抗逆转录病毒疗法（HAART）积极治疗HIV显著改善了HIV患者的健康和预后，降低了全身HIV病毒载量，主要通过增加CD 4 + T淋巴细胞恢复免疫功能。这导致了艾滋病毒相关的机会性口腔疾病，包括OPC和（OHL）的发病率大幅下降。与此形成鲜明对比的是，据报道，自从HAART广泛使用以来，口腔乳头状瘤（疣）的发病率有所增加。初步分析已经确定HPV-32是新奥尔良HIV+队列中口腔疣的主要基因型。据认为，无症状的口腔HPV感染经常发生，但口腔疣很少发生，可能是由于病毒的免疫控制。免疫反应的关键方面，防止从无症状的HPV感染的HPV疾病的进展是未知的，但以前的研究集中在HPV基因型特异性反应的主要衣壳蛋白，LI。在HIV+患者中观察到的HPV相关口腔病理学的增加率强调了对口腔HPV感染的免疫反应进行更深入了解的必要性。此外，口腔的可及性提供了一个独特的机会来进行HPV感染，免疫和发病机制的严格分析。为了启动这些研究，需要开发对口腔HPV基因型（如HPV-32）具有特异性的免疫学检测方法。我们假设，HPV-32特异性体液和细胞免疫测定可以开发和利用，以筛选人群的HPV-32特异性免疫反应。我们建议开发一种酶联免疫吸附试验（ELISA）检测血清中的HPV-32特异性抗体，淋巴细胞增殖和流式细胞术检测HPV-32特异性T细胞反应。将检测这些试验的最佳试验条件、专属性和重现性。最终，这些检测将用于研究免疫在口腔HPV感染的获得和随后的清除或进展中的作用，特别是在高度易感的HIV+患者中。 该项目的目标是开发必要的工具，以更好地了解口腔HPV感染的体液和细胞免疫反应，特别是在HIV+个体中。了解口腔HPV感染的免疫反应可能有助于预测HPV疾病的进展，以及阐明HPV相关疾病的治疗和预防策略。