A High Throughput Whole Blood Assay for Analysis of Multiple Antigen-Specific T Cell Responses in Human Mycobacterium tuberculosis Infection
In: The Journal of Immunology, Jg. 200 (2018-04-15), Heft 8
academicJournal
- 3008 - 3019
Zugriff:
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-specific T cells correlate with M. tuberculosis infection outcome in humans. To facilitate evaluation of human M. tuberculosis-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 M. tuberculosis 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 M. tuberculosis-unexposed 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-specific T cell response spectrum assay further enhances the immunological toolkit available for evaluating M. tuberculosis-specific 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-specific T cell responses associated with M. tuberculosis infection outcomes.
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A High Throughput Whole Blood Assay for Analysis of Multiple Antigen-Specific T Cell Responses in Human Mycobacterium tuberculosis Infection
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Autor/in / Beteiligte Person: | 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 |
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Zeitschrift: | The Journal of Immunology, Jg. 200 (2018-04-15), Heft 8 |
Veröffentlichung: | eScholarship, University of California, 2018 |
Medientyp: | academicJournal |
Umfang: | 3008 - 3019 |
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