Immunology Core

免疫学核心(核心D)是在孢子内部和外部咨询委员会讨论后建立的新核心，并回应了之前对我们续签申请的批评。CORE的主要目标是建立对孢子计划的四个项目正在开发和测试的疫苗的细胞和体液免疫反应的标准化分析。 通过使用统一的试剂和规程、训练有素的人员和严格的质量控制措施，将实现标准化。这些检测的集中执行将有助于对使用相同主要免疫结果衡量标准的疫苗进行面对面的比较，并提供资源的有效利用。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对免疫原性进行评估也将缓解人们对可能存在利益冲突的担忧。核心为个别项目提供的服务如下。 项目I包括血清特异性HPV16L1病毒样颗粒(VLP)酶联免疫吸附试验(EL ISA)和HPV16、31、33体外假病毒粒子中和试验。纳入后两种类型是为了评估遗传相关类型可能的交叉中和。对于项目II，中心将进行针对疫苗原、HPVL2 11-200x3蛋白的血清免疫球蛋白检测，以及针对HPV6、11和15种高危HPV类型的体外假病毒中和试验。对于接种Gardasil疫苗的亚组受试者，血清样本将在HPV16VLP ELISA法和HPV6、11、16、18、31、33和45假病毒粒子中和试验中进行检测。对于项目III和IV，Core将在用E6或E7多肽池进行过夜刺激后，对未分离的外周血单核细胞(PBMC)进行干扰素-伽马ELISPOT分析。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。二次免疫分析、针对单个项目的分析和新的免疫分析开发将由研究人员在各个项目内进行和完善，只有在完全优化和标准化后才会转移到免疫学核心。免疫学核心将与个人广泛互动 组织/病理学核心(核心C)将提供标本，生物统计/数据管理核心(核心B)将进行数据分析。

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.