Immunology/Multiplexed

获得高通量化验技术对于及时完成检验三个科学项目中所包含的假设所需的大量化验至关重要。因此，高吞吐量多路核心设备(核心C)的主要目标是在RITM安装设计流畅的装配线式工作流程、条形码样品识别、高速液体移液和多路样品分析等使能技术，这些技术将直接影响TMRC应用的所有三个科学项目： 对于项目1，多路核心设施将快速测量卡拉鲍疫苗接种的二次结果，包括Sj97抗原和同型特异性抗体水平(Ig G和Ig E)以及动物PBMC中抗原诱导的细胞因子水平(1L4、IL 5和IL-13)。对于项目2，多路核心设施将快速分析NIH UO1A1066050随机对照试验项目中母亲所生的所有N=420个PBMC，并评估Th2细胞因子(1L4、1L5和1L3)对血吸虫抗原交换和SEA以及重组Sj22和Sj97的反应水平。针对SWAP、SEA、Sj22和Sj97的特定抗体亚型也将进行分析。促炎症细胞因子(肿瘤坏死因子-a、IL-6和干扰素-γ)和炎症相关发病率，如贫血的铁缺乏标志物，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，也将被检测。在项目3中，多路核心设备将快速检测14+/-2周和32+/-2周的孕妇血浆中的TIMP-1和Tenascin C(以及FibroPlex v2中包含的另外36种胶原代谢标记物)，这些孕妇来自参加NIH UO1A1066050吡喹酮随机对照试验的母亲。我们还将在体外检测分化为合体滋养层细胞并在培养液或SEA刺激下的TIMP-1(和其他37种FibroPlex v2分析物)的水平。对于项目1、2和3，核心设施将对所有化验实施质量控制战略，包括多重化验。

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.