缺氧生产条件下功能性 CD19 CAR-T 细胞的制备

We performed in vitro phenotypic and functional assessments of CD19-directed CAR T cells produced in either 21% ( Nor CAR) or 2% ( Hyp CAR) O 2 derived from healthy donors (HDs) or patients with CLL.

Production of HD-derived CAR T cells in 2% O 2 promoted the enrichment of a na ve-like subset. Hyp CAR and Nor CAR cells were functionally distinct; CD4+ Hyp CAR cells produced more IL-2 and tumor necrosis factor than CD4+ Nor CAR cells. Production in 2% O 2 was not detrimental to viability or proliferation upon cognate antigen-stimulation and led to increased activation. After chronic stimulation in hypoxia, Hyp CAR-product remained enriched in na ve-like cells, and demonstrated cytotoxic and cytokine production capacity. In CAR T cells derived from patients with CLL, Nor CAR and Hyp CAR subsets were functionally and phenotypically comparable, but displayed different mitochondrial metabolism. DISCUSSION: We demonstrated that production in 2% O 2 is not detrimental, confers subtle but lasting functional and phenotypic changes in CAR T cells warranting further research on the impact of hypoxic production on CAR T cell functionality in hypoxic tumor microenvironments.