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Whatney, Wendy E; Gandhi, Neel R; Lindestam Arlehamn, Cecilia S; Nizam, Azhar; Wu, Hao; Quezada, Melanie J; Campbell, Angela; Allana, Salim; Kabongo, Mbuyi Madeleine; Khayumbi, Jeremiah; Muchiri, Benson; Ongalo, Joshua; Tonui, Joan; Sasser, Loren E; Fergus, Tawania J; Ouma, Gregory Sadat; Ouma, Samuel Gurrion; Beck, Allison A; Mulligan, Mark J; Oladele, Alawode; Kaushal, Deepak; Cain, Kevin P; Waller, Lance; Blumberg, Henry M; Altman, John D; Ernst, Joel D; Rengarajan, Jyothi; Day, Cheryl L
The Journal of immunology (1950), 04/2018, Volume: 200, Issue: 8Journal Article
Antigen-specific CD4 and CD8 T cells are important components of the immune response to , yet little information is currently known regarding how the breadth, specificity, phenotype, and function of -specific T cells correlate with infection outcome in humans. To facilitate evaluation of human -specific T cell responses targeting multiple different Ags, we sought to develop a high throughput and reproducible T cell response spectrum assay requiring low blood sample volumes. We describe here the optimization and standardization of a microtiter plate-based, diluted whole blood stimulation assay utilizing overlapping peptide pools corresponding to a functionally diverse panel of 60 Ags. Using IFN-γ production as a readout of Ag specificity, the assay can be conducted using 50 μl of blood per test condition and can be expanded to accommodate additional Ags. We evaluated the intra- and interassay variability, and implemented testing of the assay in diverse cohorts of -unexposed healthy adults, foreign-born adults with latent infection residing in the United States, and tuberculosis household contacts with latent infection in a tuberculosis-endemic setting in Kenya. The -specific T cell response spectrum assay further enhances the immunological toolkit available for evaluating -specific T cell responses across different states of infection, and can be readily implemented in resource-limited settings. Moreover, application of the assay to longitudinal cohorts will facilitate evaluation of treatment- or vaccine-induced changes in the breadth and specificity of Ag-specific T cell responses, as well as identification of -specific T cell responses associated with infection outcomes.
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