{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T08:38:41Z","timestamp":1774773521021,"version":"3.50.1"},"reference-count":36,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2018,4,1]],"date-time":"2018-04-01T00:00:00Z","timestamp":1522540800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,4,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Antigen-specific CD4 and CD8 T cells are important components of the immune response to Mycobacterium tuberculosis, yet little information is currently known regarding how the breadth, specificity, phenotype, and function of M. tuberculosis\u2013specific T cells correlate with M. tuberculosis infection outcome in humans. To facilitate evaluation of human M. tuberculosis\u2013specific 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 M. tuberculosis Ags. Using IFN-\u03b3 production as a readout of Ag specificity, the assay can be conducted using 50 \u03bcl 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 M. tuberculosis\u2013unexposed healthy adults, foreign-born adults with latent M. tuberculosis infection residing in the United States, and tuberculosis household contacts with latent M. tuberculosis infection in a tuberculosis-endemic setting in Kenya. The M. tuberculosis\u2013specific T cell response spectrum assay further enhances the immunological toolkit available for evaluating M. tuberculosis\u2013specific T cell responses across different states of M. tuberculosis 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 M. tuberculosis\u2013specific T cell responses associated with M. tuberculosis infection outcomes.<\/jats:p>","DOI":"10.4049\/jimmunol.1701737","type":"journal-article","created":{"date-parts":[[2018,3,14]],"date-time":"2018-03-14T16:10:15Z","timestamp":1521043815000},"page":"3008-3019","source":"Crossref","is-referenced-by-count":14,"title":["A High Throughput Whole Blood Assay for Analysis of Multiple Antigen-Specific T Cell Responses in Human <i>Mycobacterium tuberculosis<\/i> Infection"],"prefix":"10.1093","volume":"200","author":[{"name":"TBRU-ASTRa Study Group","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wendy E","family":"Whatney","sequence":"additional","affiliation":[{"name":"Emory Vaccine Center, Emory University School of Medicine , Atlanta, GA 30329"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neel R","family":"Gandhi","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Rollins School of Public Health, Emory University , Atlanta, GA 30322"},{"name":"Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cecilia S","family":"Lindestam Arlehamn","sequence":"additional","affiliation":[{"name":"Division of Vaccine Discovery, La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Azhar","family":"Nizam","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, Rollins School of Public Health, Emory University , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hao","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, Rollins School of Public Health, Emory University , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2057-3477","authenticated-orcid":false,"given":"Melanie J","family":"Quezada","sequence":"additional","affiliation":[{"name":"Emory Vaccine Center, Emory University School of Medicine , Atlanta, GA 30329"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3990-3656","authenticated-orcid":false,"given":"Angela","family":"Campbell","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Rollins School of Public Health, Emory University , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salim","family":"Allana","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Rollins School of Public Health, Emory University , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5158-6860","authenticated-orcid":false,"given":"Mbuyi Madeleine","family":"Kabongo","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, Rollins School of Public Health, Emory University , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeremiah","family":"Khayumbi","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benson","family":"Muchiri","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4968-6176","authenticated-orcid":false,"given":"Joshua","family":"Ongalo","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joan","family":"Tonui","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3348-9933","authenticated-orcid":false,"given":"Loren E","family":"Sasser","sequence":"additional","affiliation":[{"name":"Emory Vaccine Center, Emory University School of Medicine , Atlanta, GA 30329"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tawania J","family":"Fergus","sequence":"additional","affiliation":[{"name":"Department of Medicine, New York University School of Medicine , New York, NY 10016"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory Sadat","family":"Ouma","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samuel Gurrion","family":"Ouma","sequence":"additional","affiliation":[{"name":"Center for Global Health Research, Kenya Medical Research Institute , Kisumu 40100,","place":["Kenya"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Allison A","family":"Beck","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mark J","family":"Mulligan","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Emory University School of Medicine , Atlanta, GA 30322"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alawode","family":"Oladele","sequence":"additional","affiliation":[{"name":"DeKalb County Board of Health , Atlanta, GA 30030"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3521-1257","authenticated-orcid":false,"given":"Deepak","family":"Kaushal","sequence":"additional","affiliation":[{"name":"Tulane National Primate Research Center , Covington, LA 70433"},{"name":"Department of Microbiology and Immunology, Tulane Health Sciences Center , New Orleans, LA 70112"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin P","family":"Cain","sequence":"additional","affiliation":[{"name":"Division of Global HIV and Tuberculosis, U.S. Centers for Disease Control and Prevention , Kisumu 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Vaccine Immunol."},{"key":"2025030610143753300_r16","doi-asserted-by":"crossref","first-page":"e74080","DOI":"10.1371\/journal.pone.0074080","article-title":"Analysis of host responses to Mycobacterium tuberculosis antigens in a multi-site study of subjects with different TB and HIV infection states in sub-Saharan Africa","volume":"8","author":"GCGH Biomarkers for TB consortium","year":"2013","journal-title":"PLoS One"},{"key":"2025030610143753300_r17","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1111\/imr.12274","article-title":"T cells and adaptive immunity to Mycobacterium tuberculosis in humans","volume":"264","author":"Jasenosky","year":"2015","journal-title":"Immunol. Rev."},{"key":"2025030610143753300_r18","doi-asserted-by":"crossref","DOI":"10.1128\/microbiolspec.TBTB2-0016-2016","article-title":"Human immunology of tuberculosis","author":"Scriba","year":"2017","journal-title":"Microbiol. 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