Glioblastoma-targeted CD4+ CAR T cells mediate superior antitumor activity

Chimeric antigen receptor-modified (CAR-modified) T cells have shown promising therapeutic effects for hematological malignancies, yet limited and inconsistent efficacy against solid tumors. The refinement of CAR therapy requires an understanding of the optimal characteristics of the cellular products, including the appropriate composition of CD4(+) and CD8(+) subsets. Here, we investigated the differential antitumor effect of CD4(+) and CD8(+) CAR T cells targeting glioblastomaassociated (GBM-associated) antigen IL-13 receptor alpha 2 (IL13R alpha 2). Upon stimulation with IL13R alpha 2(+) GBM cells, the CD8(+) CAR T cells exhibited robust short-term effector function but became rapidly exhausted. By comparison, the CD4(+) CAR T cells persisted after tumor challenge and sustained their effector potency. Mixing with CD4(+) CAR T cells failed to ameliorate the effector dysfunction of CD8(+) CAR T cells, while surprisingly, CD4(+) CAR T cell effector potency was impaired when coapplied with CD8(+) T cells. In orthotopic GBM models, CD4(+) outperformed CD8(+) CAR T cells, especially for long-term antitumor response. Further, maintenance of the CD4(+) subset was positively correlated with the recursive killing ability of CAR T cell products derived from GBM patients. These findings identify CD4(+) CAR T cells as a highly potent and clinically important T cell subset for effective CAR therapy.