Immunology Core

免疫学核心

基本信息

批准号：
7727559
负责人：
Chien-Fu Hung
金额：
$ 18.41万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7727559
关键词：
Advisory Committees Biological Assay Biometry CD8B1 gene Cellular Assay Cellular Immunity Clinical Trials Conflict of Interest Critiques Data Data Analyses Data Storage and Retrieval Development Ensure Enzyme-Linked Immunosorbent Assay Flow Cytometry Funding Future Gardasil Goals HPV-High Risk Head Human Papillomavirus Human Resources Human papilloma virus infection Human papillomavirus 16 Human papillomavirus 6 Immune response Immunoglobulin G Immunology Immunology procedure In Vitro Individual Instruction Interferon Type II Label Laboratories Lymphocyte Malignant neoplasm of cervix uteri Measures Outcome Measure Pathology Peptides Performance Peripheral Blood Mononuclear Cell Phase Phenotype Prevention Procedures Proteins Protocols documentation Quality Control Reagent Reproduction spores Research Personnel Resources Sampling Serum Services Specimen Staining method Stains Standardization Testing Tissues Training Vaccinated Vaccine Antigen Vaccines Virus-like particle Work assay development base cytokine data acquisition data management database structure design enzyme linked immunospot assay good laboratory practice immunogenicity interest novel novel vaccines primary outcome programs response

项目摘要

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.

免疫学核心（核心D）是孢子讨论后建立的新核心内部和外部咨询委员会以及对我们更新的先前批评应用。核心的主要目标是建立细胞和体液的标准化测定对孢子计划的四个项目开发和测试的疫苗的免疫反应。标准化将通过使用统一的试剂和协议，训练有素的人员和严格的质量控制措施。这些测定的集中性能将有助于正面比较使用相同主要免疫结局指标的疫苗，并有效地使用资源。因为一些孢子调查人员对正在疫苗有财务利益经过测试，通过独立指示实验室评估免疫原性也将使人们对可能的利益冲突。核心为各个项目提供的服务如下。对于项目I，测定包括血清IgG特异性HPV 16 L1病毒（如颗粒（VLP）酶）免疫吸附测定（ELISA）和HPV 16、31和33体外伪动物中和测定。后者包括2种类型以评估遗传相关类型的可能的交叉中和。对于第二个项目，核心将执行ELISA检测针对疫苗的血清IgG，HPV L2 11-200x3蛋白以及对HPV 6和11和15高风险HPV类型的体外假期中和测定。对于子集在用Gardasil接种疫苗的受试者中，将在HPV 16 VLP ELISA和HPV 6、11中测试血清样品 16、18、31、33和45个假膜中和测定。对于III和IV项目，核心将执行一夜之间，对未分流的外周血单核细胞（PBMC）进行了伽马ELISPOT测定用E6或E7肽池刺激。反应淋巴细胞的CD8 +和CD4 +表型细胞内细胞因子染色和流式细胞术将证实。次生免疫学测定，将对单个项目和新型免疫学分析开发进行特定的测定法，并将进行由各个项目中的调查人员完善，仅转移到免疫学核心完全优化和标准化时。免疫学核心将与个体广泛相互作用项目/病理核心（核心C），将提供标本，以及 BioStatistLCS/数据管理核心（Core B），将执行数据分析。相关性（请参阅说明）：免疫学核心将进行标准化测定，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。