Immunology/Multiplexed

免疫学/多重

• 批准号: 8716657
• 负责人: Luz Acosta
• 金额: $ 5.39万
• 依托单位: RESEARCH INSTITUTE FOR TROPICAL MEDICINE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8716657
• 关键词: American; Anemia; Animals; Antibodies; Antigens; Bar Codes; Biological Assay; Boxing; Buffaloes; Child; Clinical; Collagen; Core Facility; Data; Databases; Development; Documentation; Enrollment; Equipment and Supplies; Erythropoietin; Ferritin; Fibrosis; Funding; Goals; Hematology; Housing; IL5 gene; IgE; Immunoglobulin G; Immunology; In Vitro; Inflammatory; Instruction; Interferons; Interleukin-13; Iron; Laboratories; Leptin; Life Cycle Stages; Liquid substance; Location; Maintenance; Measures; Metabolism; Methods; Morbidity - disease rate; Mothers; Pathologist; Plasma; Praziquantel; Pregnancy; Procedures; Quality Control; Reading; Reagent; Recombinants; Robot; Running; Sampling; Scanning; Schistosoma; Schistosomiasis; Serum; Services; Speed; System; TNF gene; Technology; Tenascin; Testing; Tissue Inhibitor of Metalloproteinase-1; Training; Transferrin Receptor; United States National Institutes of Health; Vaccination; Work; assay development; base; college; cost; cost effective; cytokine; design; high throughput screening; instrument; iron deficiency; operation; programs; response; secondary outcome; synthetic polymer Bioplex; trophoblast; vaccine development

Access to high throughput assay technology is essential for the timely completion of the large quantity of assays required to test the hypotheses contained in the three scientific projects. It is therefore the primary aim of a high throughput Multiplex Core Facility (Core C) to install at RITM a smoothly designed, assembly line like work flow, the enabling technologies of barcode sample identification, high speed liquid pipetting and multiplexed sample analysis that will directly impact all three of the Scientific Projects for this TMRC application: For Project 1, the Mulitplex Core facility will rapidly measure secondary outcomes of vaccination in carabaos that will include Sj97 antigen and isotype specific antibody levels (IgG and IgE) and antigen induced cytokine levels from the animal's PBMCs (1L4, IL5 and IL-13). For Project 2, the Multiplex Core facility will rapidly analyze all N=420 PBMCs from children born to mothers included in the NIH UO1A1066050 RCT project and assess levels of Th2 cytokines (1L4,1L5, and 1L3) in response to both schistosome antigens SWAP and SEA, and recombinant Sj22 and Sj97. Specific antibody isotypes will be also be analyzed targeting SWAP, SEA, Sj22 and Sj97. Pro-inflammatory cytokines (TNF-a, lL-6, and IFN-y) and inflammatory associated morbidity such as Iron deficiency markers of anemia such as Hgb, Leptin, Ferritin, Erythropoietin, and Soluble Transferrin Receptor feritin will likewise be measured. For Project 3, the Multiplex Core facility will rapidly measure TIMP-1 and Tenascin C (and 36 additional markers of collagen metabolism contained in FibroPlex v2) in maternal plasma collected at 14 +/- 2 wks and 32 +/- 2 wks gestation from mothers enrolled in the NIH UO1A1066050 Praziquantel RCT. We will also measure TIMP-1 (and 37 other FibroPlex v2 analytes) levels in isolated term trophoblasts differentiated into syntytiotrophoblasts and stimulated with media or SEA in vitro. For Projects 1, 2, and 3, the Core Facility will implement a Quality Control strategy for all assays, including multiplex assays.

获得高通量分析技术对于及时完成测试三个科学项目中包含的假设所需的大量分析至关重要。因此，高通量多路复用核心设施（核心C）的主要目标是在RITM安装一个设计平稳的装配线，如工作流程，条形码样品识别，高速液体移液和多路复用样品分析的使能技术，这将直接影响TMRC应用的所有三个科学项目： 对于项目1，Mulitplex Core设施将快速测量Carabao中疫苗接种的次要结果，包括Sj 97抗原和同种型特异性抗体水平（IgG和IgE）以及动物PBMC中抗原诱导的细胞因子水平（1 L4、IL-5和IL-13）。对于项目2，Multiplex Core设施将快速分析NIH UO 1A 1066050随机对照试验项目中母亲所生儿童的所有N=420个PBMC，并评估针对血吸虫抗原SWAP和SEA以及重组Sj 22和Sj 97的Th 2细胞因子（1 L 4、1 L 5和1 L 3）水平。还将针对SWAP、SEA、Sj 22和Sj 97分析特异性抗体同种型。同样将测量促炎细胞因子（TNF-α、IL-6和IFN-γ）和炎症相关的发病率，例如贫血的缺铁标志物，例如Hgb、瘦素、铁蛋白、促红细胞生成素和可溶性转铁蛋白受体铁蛋白。对于项目3，Multiplex Core机构将快速测量从入组NIH UO 1A 1066050吡喹酮RCT的母亲中在妊娠14 +/- 2周和32 +/- 2周采集的母体血浆中的TIMP-1和腱生蛋白C（以及Fibroblastin v2中包含的36种额外的胶原代谢标志物）。我们还将测量在分化成合体滋养层并在体外用培养基或SEA刺激的分离的足月滋养层中的TIMP-1（和37种其他Fibroblast v2分析物）水平。对于项目1、2和3，核心机构将对所有测定（包括多重测定）实施质量控制策略。