Lentivirus vector construction and lentivirus production were established as previously described (Hu et al., 2021 (link)). To generate TCR-transduced-CD4+ T and TCR-transduced-CD4+ Jurkat cells, CD4+ T cells were stimulated with 1 μg/ml OKT3 antibody and 1 μg/ml anti-CD28 antibody (Miltenyi Biotec) and cultured in a complete medium containing 100 IU/ml IL-2 for 48 h. CD4+ Jurkat cells and stimulated CD4+ T cells were seeded into 24-well-plates and incubated with 500 μl lentivirus and 500 μl RPMI 1640 for 6 h. Next, 500 μl complete medium was added to each well. After 24 h, the original medium was replaced with a 2-ml complete medium and incubated for 48 h. Then, the cells were harvested and used for subsequent assays.
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