The role of the human immune monitoring core (HIMC, Core C) will be to (1) provide standardized, state-ofthe art Immune monitoring assays at the RNA, protein, and cellular level, to support research projects within the U19;(2) to test and develop new technologies for immune monitoring that may be useful In the context of the U19;and (3) to efficiently archive, report, and mine data from immune monitoring studies, so as to increase the value of the data and to assist In biomarker discovery.
Specific Aim 1 : Standardized, state-of-the-art immune monitoring assays. The HIMC has validated a set of Immune monitoring assays that will be available to U19 research projects. These include genome-wide RNA microarrays, multiplex Luminex cytokine assays, immunophenotyping, phosphoepitope flow cytometry, CFSE dye dilution assays for proliferation, and intracellular cytokine staining. An ELISPOT reader is also available for readout of ELISPOT assays.
Specific Aim 2 : New technology for immune monitoring. The HIMC is evaluating multiple new platforms with potential for immune monitoring, including: isoelectric focusing analysis of phosphoproteins (CellBiosciences);chemiluminescent cytokine detection (MesoScale Discovery);biomolecular interaction analysis (ForteBlo);multiplexed tetramer analysis;flow cytometry with time-of-flight mass spectrometry (CyTof);qPCR arrays on sorted cell populations (Fluidigm Blomark);and specialized microarrays for Immunologically relevant genes and for pathogen detection (Agilent).
Specific Aim 3 : Databaslng. The HIMC is Implementing the use of collaborative online databases for flow cytometry, Luminex, and microarray data, and is also evaluating data aggregation and mining programs such as Tibco Spotfire.
The use of standardized assays and data management will facilitate mining across studies and greatly Increase the value of the data for discovery of new biomarkers of vaccine efficacy and disease protection. New technology developed may create opportunities to discover such biomarkers In ways not possible before.
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