Characteristics of GM-CSF–secreting CD4+ T cells.

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Characteristics of GM-CSF–secreting CD4+ T cells. Characteristics of GM-CSF–secreting CD4+ T cells. (A) Splenocytes from mice infected with HSV-2 cutaneously or intravaginally or infected with M. tuberculosis by aerosol route underwent stimulation with UV-inactivated HSV-2 or ESAT-6 peptide, respectively, and intracellular cytokine staining. The cells shown are single cells from the lymphocyte gate (FSC/SSC) that are positive for CD4 and CD44 and negative for LIVE/DEAD stain, CD8α, B220, and MHC II. The figure represents results of two independent experiments. (B) Splenocytes from mice infected with HSV-2 cutaneously were processed as in (A). Pie chart depicts the fraction of GM-CSF–secreting Ag-experienced CD4+ T cells that also secrete cytokines that define multifunctionality. Each shade represents number(s) of additional cytokine(s) secreted. The figure represents results of two independent experiments. (C) Splenocytes from mice infected with HSV-2 cutaneously or i.v. underwent stimulation with UV-inactivated HSV-2 and intracellular cytokine staining. Contour plots depict GM-CSF–secreting CD4+ T cells obtained postinfection through indicated routes. Bar graphs show the percentage of CD4+ T cells that secrete GM-CSF. The figure represents results of two independent experiments. *p < 0.05 for comparisons between i.v. and skin (SK) (Student t test). (D) Splenocytes from OT-II TCR-Tg mice were cultured in the presence of OVA peptide (OVA323–339) and cytokines/Abs as indicated to generate Th1 cells (left) or predominantly IL-3–producing Th cells (right). The contour plot represents coexpression of GM-CSF with IL-3 with the numbers representing the percentage of CD4+ T cells for each of the corresponding gates. The figure represents results of two independent experiments. Shajo Kunnath-Velayudhan et al. ImmunoHorizons 2019;3:161-171 Copyright © 2019 The Authors