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A high throughput whole blood assay for analysis of multiple antigen-specific T cell responses in human Mycobacterium tuberculosis infection1
Ist Teil von
The Journal of immunology (1950), 2018-03, Vol.200 (8), p.3008-3019
Erscheinungsjahr
2018
Link zum Volltext
Quelle
EZB Electronic Journals Library
Beschreibungen/Notizen
Antigen (Ag)-specific CD4 and CD8 T cells are important components of the
immune response to
Mycobacterium tuberculosis
(Mtb), yet little
information is currently known regarding how the breadth, specificity, phenotype
and function of Mtb-specific T cells correlate with Mtb infection outcome in
humans. To facilitate evaluation of human Mtb-specific T cell responses
targeting multiple different Ags, we sought to develop a high throughput and
reproducible T cell response spectrum assay (RSA) 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 Mtb Ags. Using
IFN-γ production as a readout of Ag specificity, the assay can be
conducted using 50µl of blood per test condition and can be expanded to
accommodate additional Ags. We evaluated the intra- and inter-assay variability,
and implemented testing of the assay in diverse cohorts of Mtb-unexposed healthy
adults, foreign-born adults with latent Mtb infection (LTBI) residing in the
U.S., and TB household contacts with LTBI in a TB-endemic setting in Kenya. The
Mtb-specific T cell RSA further enhances the immunological toolkit available for
evaluating Mtb-specific T cell responses across different states of Mtb
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
Mtb-specific T cell responses associated with Mtb infection outcomes.