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Characterization of lung infection-induced TCRγδ T cell phenotypes by CyTOF mass cytometry

Wanke-Jellinek, L., Keegan, J.W., Dolan, J.W., Lederer, J.A.

T cell receptor γδ cells are known to be the primary effector T cells involved in the response to bacterial infections, yet their phenotypic characteristics are not as well established as other T cell subsets. In this study, we used cytometry by time-of-flight mass cytometry to better characterize the phenotypic response of T cell receptor γδ cells to Streptococcus pneumoniae lung infection. Mice were infected, and cells from lung washouts, spleen, and lymph nodes were stained to detect cell-surface, intracellular, and signaling markers. We observed that infection caused a significant increase in T cell receptor γδ cells, which expressed high interferon-γ and interleukin-17A levels. Profiling T cell receptor γδ cells by cytometry by time-of-flight revealed that activated γδ T cells uniquely coexpressed cell-surface Gr-1, cluster of differentiation 14, and cluster of differentiation 274 (programmed death-ligand 1). Further classification of Gr-1 expression patterns on T cell receptor γδ cells demonstrated that Gr-1+ T cell receptor γδ cells were the primary source of interferon-γ, whereas Gr-1- cells mostly expressed interleukin-17A. Gr-1+ T cell receptor γδ cells also showed higher ζ-chain-associated protein kinase 70, p38, and 4eBP1 signaling in response to infection as compared with Gr-1- T cell receptor γδ cells. Taken together, Gr-1 expression patterns on γδ T cells in the lung provide a robust marker to differentiate interferon-γ- and interleukin-17A-producing subsets involved in the early immune response to bacterial pneumonia.

Citation

Wanke-Jellinek, L., Keegan, J.W., Dolan, J.W., Lederer, J.A. "Characterization of lung infection-induced TCRγδ T cell phenotypes by CyTOF mass cytometry" Journal of Leukocyte Biology (2015): 483–93