Generation of HVS-transformed human primary T cells and cytotoxicity

PMBCs from healthy donors or patients with XLP1 were stimulated in vitro with 20 μg/mL PHA and then cultured in X-VIVO media with 5% human serum for a period of 72 hours. After 72 hours, IL-2 (20 ng/mL) was added to supplement media, and cells were incubated for a further 48 hours. HVS viral supernatant generated by using owl monkey kidney cells, as previously published, was added to cultured cells seeded at a density of 2.0 × 10⁶ cells/well of a 48-well plate. The phenotype of transformed cells was assessed at 2-week intervals to determine cell longevity and viability.