Supplementary Figure 4 : Defective naive CD4 T cell proliferation in patient A-I, ELISA analysis of A-I and family B patients, and glutamine/cytokines rescue of IFN-γ in the family D patients.

From: Germline hypomorphic CARD11 mutations in severe atopic disease

Supplementary Figure 4

(a) Blastogenesis and proliferation of isolated naïve CD4+ cells from representative healthy control (HC) vs. CARD11 patients after anti-CD3/CD28 activation for 5 days. CellTrace Violet (Violet) staining was used for tracking the number of the cell divisions. (b) Culture media from the PMA/ionomycin-treated patients’ PBMCs for 6 h were collected, and the IFNγ, IL-4, IL-13 and IL-5 secretions were measured by ProcartaPlex ELISA (eBioscience) (mean ± s.e.m.). (c) Intracellular flow cytometry identifying IFNγ/IL-4-producing cell ratio from CD3+CD8 CD45RO+ T cells within PBMC from CARD11 patients of family D and travel control (TC). PBMCs were cultured with anti-CD3/CD28 activation for 5 days, with or without cytokines (Th0) and plus 3 mM glutamine.