CD4+ T cells were immunomagnetically isolated from the spleens of wild-type mice. For induction of Th1 differentiation, we cultured CD4+ T cells with anti-CD3/CD28 antibody (Dynabeads Mouse T activator, Life Technologies, Carlsbad, CA), IL-12 (10 ng/ml, R&D Systems) and anti-IL-4 (10 µg/ml, Bio X Cell) for 72 h. For induction of Th17 differentiation, we cultured CD4+ T cells with anti-CD3/CD28 antibody (Dynabeads Mouse T activator), IL-6 (10 ng/ml, R&D Systems), TGF-β (2 ng/ml, R&D Systems), anti-IL-2(10 µg/ml, Bio X Cell), anti-IL-4 (10 µg/ml, Bio X Cell) and anti-IFN-γ (10 µg/ml, R&D Systems) for 72 h. For induction of Treg differentiation, we cultured CD4+ T cells with anti-CD3/CD28 antibody (Dynabeads Mouse T activator), TGF-β (5 ng/ml, R&D Systems), IL-2 (10 ng/ml, Bio X Cell), anti-IL-4 (10 µg/ml, Bio X Cell) and anti-IFN-γ (10 µg/ml, R&D Systems) for 72 h. Flow cytometry was used for the differentiation and activation of CD4+ T cells.
Su Y., Sun X., Liu X., Qu Q., Yang L., Chen Q., Liu F., Li Y., Wang Q., Huang B., Huang X.H, & Zhang X.J. (2022). hUC-EVs-ATO reduce the severity of acute GVHD by resetting inflammatory macrophages toward the M2 phenotype. Journal of Hematology & Oncology, 15, 99.
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