Human cd4 t cell isolation kit

HEK293T and THP-1 cells were obtained from ATCC and cultured in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 1 U/ml penicillin and 100 mg/ml streptomycin (Gibco). Human CD14 + cells were isolated from healthy donor PBMCs using MojoSort Human CD14 Selection Kit (Biolegend #480026), and were cultured with 50 ng/ml recombinant human GM-CSF (BioLegend #572904) and 50 ng/ml recombinant human M-CSF (BioLegend #574806) for 6 days to obtain monocytes derived macrophages. CD4 + T cells from human tonsil biopsies were purified by sorting. Blood CD4 + T cells were isolated from healthy donor or patient PBMCs using a human CD4 + T cell isolation kit (BioLegend #480010). Purified CD4 + T cells were used without stimulation, or co-stimulated with plate-bound CD3 (Biolegend #300333) and soluble CD28 (Biolegend #302943) antibodies with the presence of 20 ng/μl IL-2 (Biolegend #589106) for 3 days.