Immunology/Multiplexed

获得高通量分析技术对于及时完成大量检测至关重要

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.