Immunology Core

免疫学核心(核心D)是孢子经过讨论后建立的新核心 内部和外部咨询委员会，并回应先前对我们续签的批评 申请。核心的主要目标是建立细胞和体液的标准化分析 孢子计划的四个项目正在开发和测试疫苗的免疫反应。 将通过使用统一的试剂和方案、训练有素的人员和严格的 质量控制措施。这些化验的集中执行将有助于面对面的比较 使用相同的主要免疫结果指标并提供有效使用的疫苗 资源。因为一些孢子调查人员对正在开发的疫苗有经济利益 经过测试，由独立指导的实验室进行免疫原性评估也将缓解人们对 可能存在利益冲突。核心为个别项目提供的服务如下。 对于项目I，检测包括血清特异性HPV16L1病毒样颗粒(VLP)酶联检测 免疫吸附试验(ELISA法)和HPV16、31、33体外假病毒中和试验。后者 包括两种类型，以评估遗传相关类型可能的交叉中和。就项目II而言， CORE将进行酶联免疫吸附试验，以检测针对疫苗原HPVL2 11-200x3蛋白的血清免疫球蛋白 体外假病毒中和试验检测高危型HPV6型、11型和15型。对于子集 对于接种Gardasil的受试者，血清样本将在HPV16VLP和HPV6，11， 16、18、31、33和45个假病毒粒子中和试验。对于项目III和IV，核心将执行干扰素- 过夜后外周血单个核细胞(PBMC)的Gamma ELISPOT分析 用E6或E7多肽池刺激。免疫应答淋巴细胞的CD8+和CD4+表型 将通过细胞内细胞因子染色和流式细胞术进行确认。二次免疫学检测， 将进行针对单个项目的分析和新的免疫学分析开发，并 由研究人员在个别项目中完善，并仅转移到免疫学核心 当完全优化和标准化时。免疫学核心将与个人广泛互动 项目以及将提供标本的组织/病理核心(核心C)，以及 生物统计/数据管理核心(核心B)，将进行数据分析。 相关性(请参阅说明)： 免疫学核心将进行标准化分析，以评估对新疫苗的免疫反应 预防和治疗人乳头瘤病毒感染。

The Immunology Core (Core D) is a new core that was established after discussion by the SPORE Internal and External Advisory Committees and in response to the previous critique of our renewal application. The principal goal of the Core is to establish standardized assays of cellular and humoral immune responses to the vaccines being developed and tested by the four projects of the SPORE program. Standardization will be achieved by use of uniform reagents and protocols, trained personnel and stringent quality control measures. Centralized performance of these assays will facilitate head-to-head comparisons of vaccines that use the same primary immunological outcome measures and provide for efficient use of resources. Because some SPORE investigators have a financial interest in the vaccines that are being tested, assessment of immunogenicity by an independently directed laboratory will also allay concerns over possible conflicts of interest. The services provided by the Core to the individual projects are the following. For Project I, the assays include serum IgG-specific HPV 16 L1 virus like particle (VLP) enzyme linked immunosorbent assay (ELISA) and HPV 16, 31 and 33 in vitro pseudovirion neutralization assays. The latter 2 types are included to assess possible cross neutralization of genetically related types. For Project II, the Core will perform an ELISA to detect serum IgG directed against the vaccinogen, HPV L2 11-200x3 protein and in vitro pseudovirion neutralization assays for HPV 6 and 11 and 15 high risk HPV types. For the subset of subjects vaccinated with Gardasil, serum samples will be tested in the HPV 16 VLP ELISA and HPV 6, 11, 16, 18, 31, 33 and 45 pseudovirion neutralization assays. For Projects III and IV, the Core will perform IFN- gamma ELISPOT assays on unfractionated peripheral blood mononuclear cells (PBMC) following overnight stimulation with pools of E6 or E7 peptides. The CD8+ and CD4 + phenotype of the responding lymphocytes will be confirmed by intracellular cytokine staining and flow cytometry. Secondary immunologic assays, assays specific to an individual project and novel immunologic assay development will be performed and perfected by the investigators within the individual projects and only transferred to the Immunology Core when fully optimized and standardized. The Immunology Core will interact extensively with the individual projects as well as the Tissue/Pathology Core (Core C), which will provide specimens, and the Biostatistlcs/Data Management Core (Core B), which will perform data analyses. RELEVANCE (See Instructions): The Immunology Core will perform standardized assays to assess immune responses to new vaccines for prevention and treatment of human papillomavirus infection.