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
A centralized high throughput multiplex facility is essential in the timely completion of large quantity of assays for all three scientific projects. This facility can provide a cost effective standardized tracking database and sample storage and implement a centralized quality control plan for all assays required in all three scientific projects.
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