Immunology/Multiplexed

获得高通量的检测技术是实现大批量检测的关键 测试三个科学项目中包含的假设所需的化验结果。因此，它是主要的 高吞吐量多路复用核心设备(核心C)的目标是在RITM安装设计顺畅的组件 线状工作流程、条形码样品识别、高速液体输送和条形码采集的使能技术 将直接影响本TMRC的所有三个科学项目的多重样品分析 适用范围： 对于项目1，多路核心设施将快速衡量卡拉博疫苗接种的次要结果 这将包括Sj97 Anfigen和同型特异性抗体水平(IgG和IgE)和Anfigen诱导的细胞因子 动物PBMCs(1L4、IL-5和IL-13)水平。对于项目2，多路复用核心设施将迅速 分析NIH UO1A1066050随机对照试验项目中母亲所生的所有N=420个PBMC 评估Th2细胞因子(1L4、1L5和1L3)水平对血吸虫成虫交换和SEA的反应， 和重组Sj22和Sj97。还将分析特定的抗体亚型，包括掉期、SEA、 Sj22和Sj97。促炎细胞因子(肿瘤坏死因子-0、白细胞介素6和干扰素-γ)与炎症相关发病率 例如贫血缺铁标志物，如HGB、Lepfin、铁蛋白、红细胞生成素和可溶性 转铁蛋白受体受精蛋白也将同样被测量。对于项目3，多路复用核心设施将迅速 检测TIMP-1和Tenascin C(以及FibroPlex中包含的36个额外的胶原代谢标记物 在14+/-2周和32+/-2周时从美国国立卫生研究院登记的母亲采集的孕妇血浆中 UO1A1066050吡喹酮RCT。我们还将测量TIMP-1(和其他37种FibroPlex v2分析物)的水平 在体外培养条件下，滋养层细胞分化为合体滋养层细胞，并用培养液或SEA刺激。 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 多重分析。 相关性(请参阅说明)： 一个集中的、高吞吐量的多路传输设施对于最终完成大量的 所有三个科学项目的化验结果。该设施可以提供成本效益标准化的跟踪 数据库和样品存储，并对所有所需的所有分析实施集中的质量控制计划 三个科学项目。

Access to high throughput assay technology is essenfial for the fimely complefion 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 highthroughput Mulfiplex Core Facility (Core C) to install at RITM a smoothly designed, assembly line like work fiow, the enabling technologies of barcode sample identificafion, high speed liquid pipetfing and mulfiplexed sample analysis that will direcfiy impact all three of the Scientific Projects for this TMRC applicafion: For Project 1, the Mulitplex Core facility will rapidly measure secondary outcomes of vaccinafion in carabaos that will include Sj97 anfigen and isotype specific anfibody levels (IgG and IgE) and anfigen induced cytokine levels from the animal's PBMCs (1L4, IL5 and IL-13). For Project 2, the Mulfiplex 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 anfigens SWAP and SEA, and recombinant Sj22 and Sj97. Specific anfibody isotypes will be also be analyzed targefing SWAP, SEA, Sj22 and Sj97. Pro-inflammatory cytokines (TNF-o, lL-6, and IFN-y) and inflammatory associated morbidity such as Iron deficiency markers of anemia such as Hgb, Lepfin, Ferrifin, Erythropoefin, 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 addifional markers of collagen metabolism contained in FibroPlex v2) in maternal plasma collected at 14 +/- 2 wks and 32 +/- 2 wks gestafion 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 differenfiated into synfiotrophoblasts 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 mulfiplex assays. RELEVANCE (See instructions): A centralized highthroughput mulfiplex facility is essenfial in the fimely complefion of large quantity of assays for all three scientific projects. This facility can provide a cost effecfive standardized tracking database and sample storage and implement a centralizedquality control plan for all assays required in all three scientific projects.