Immunology Core

免疫学核心（核心D）是经过SPORE内部和外部咨询委员会讨论后建立的一个新核心，并回应了之前对我们更新申请的批评。核心的主要目标是建立标准化的细胞和体液免疫反应的疫苗正在开发和测试的四个项目的孢子计划。 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来实现标准化。这些检测试剂盒的集中性能将促进使用相同主要免疫学结局指标的疫苗的头对头比较，并提供资源的有效利用。因为一些SPORE调查人员对正在进行的疫苗有经济利益， 由独立的实验室对免疫原性进行评估，也将减轻对可能的利益冲突的担忧。核心小组为各个项目提供的服务如下。 对于项目I，检测包括血清IgG特异性HPV 16 L1病毒样颗粒（VLP）酶联免疫吸附试验（ELISA）和HPV 16、31和33体外假病毒体中和试验。包括后2种类型，以评估遗传相关类型的可能交叉中和。对于项目II，中心将进行ELISA检测针对疫苗原的血清IgG、HPV L2 11- 200 x3蛋白以及HPV 6和11以及15型高危HPV的体外假病毒体中和试验。对于接种Gardasil疫苗的受试者亚组，将在HPV 16 VLP ELISA和HPV 6、11、16、18、31、33和45假病毒体中和试验中检测血清样本。对于项目III和IV，核心将在E6或E7肽池过夜刺激后对未分级外周血单核细胞（PBMC）进行IFN-γ ELISPOT测定。应答淋巴细胞的CD 8+和CD 4+表型 将通过细胞内细胞因子染色和流式细胞术确认。二级免疫测定、特定于单个项目的测定和新型免疫测定开发将由研究者在单个项目内进行和完善，只有在完全优化和标准化后才转移到免疫学核心。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）和生物统计学/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.