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. Author manuscript; available in PMC: 2022 Feb 1.
Published in final edited form as: J Allergy Clin Immunol. 2020 Jul 21;147(2):470–483. doi: 10.1016/j.jaci.2020.04.063

Figure 3. Enhanced cytokine production with stimulation in IMDM compared to RPMI.

Figure 3.

(A) Th2 cells were cultured in vitro for 3 days with plate-bound anti-CD3 (2 μg/ml), soluble anti-CD28 (2 μg/ml), human IL-2 (12.5 ng/ml), murine IL-4 (25 ng/ml), anti-IFN-γ (5 μg/ml), and anti-IL-12 (2 μg/ml). Th2 cells were re-plated on day 3 without anti-CD3 and rested for 2 additional days. On day 5 of the culture, Th2 cells were stimulated with PMA/ionomycin in complete RPMI versus IMDM. IL-4 and IL-13 intracellular staining is shown. Pre-gated on live CD4+ cells. (B) IL-4 and IL-13 intracellular staining in day 7 medLN cells from i.n. immunization with Alternaria alternata and NP-OVA. Cells were stimulated with PMA/ionomycin in complete RPMI versus IMDM. Pre-gated on Tfh cells (live CD4+ TCRβ+ CD44+ PD-1+ CXCR5+). Samples were acquired on a BD LSR II (BD Biosciences).