Rivera Reyes, A. M. | bioRxiv Science

bioRxiv2025

Neoantigens drive adoptively transferred CD8 T cells to long-lived effectors mediated by lymph node trafficking

Adoptive transfer of neoantigen-reactive T lymphocytes mediates potent responses against solid tumors in patients while often limiting toxicity to normal tissues; however, the mechanisms governing the trafficking, activation, and superior antitumor function of infused T cells remain unknown. Using a clinically relevant TCR-transgenic T cell therapy model, we examined CD8 T cell responses to melanoma expressing either wild-type or mutated antigen. Neoantigen expression conferred robust tumor regression, durable cures, and long-term protective immunity. Mechanistically, T cells reacting to neoantigen exhibited enhanced cytokine and chemokine production, heightened effector function, and sustained persistence within the blood, tumor, and lymph nodes. Notably, trafficking through secondary lymphoid organs was necessary for T cell persistence and antitumor efficacy. These findings highlight the critical role of T cell trafficking to the lymph nodes in shaping neoantigen-specific antitumor responses and offers insight for improving adoptive cellular therapies.

cancer biology↗

bioRxiv2022

Augmenting TCR signal strength and ICOS costimulation results in metabolically fit and therapeutically potent human CAR Th17 cell therapy

Adoptive cell transfer (ACT) therapy with IL-17 producing human T cells elicits potent antitumor activity in preclinical models. However, further refinement of this novel approach is needed to position it for clinical application. While activation signal strength differentially regulates IL-17 production by human CD4+ T cells, the degree to which TCR and co-stimulation signal strength impacts antitumor Th17 cell immunity remains poorly understood. We discovered that decreasing TCR/co-stimulation signal strength by incremental reduction of CD3/co-stimulation beads in a Th17 culture progressively diminished their effector memory phenotype but enhanced their polyfunctionality. Additional investigation revealed that Th17 cells stimulated with CD3/ICOS beads produced more IL-17A, IFN{gamma}, IL-2 and IL-22 than those stimulated with CD3/CD28 beads, regardless of signal strength. Th17 cells propagated with 30-fold fewer CD3/ICOS beads (weak signal strength, 1 bead per 10 T cells) were less reliant on glucose for growth compared to those stimulated with the standard, strong signal strength (3 beads per T cell). Further metabolomic analysis revealed Th17 cells weakly simulated with CD3/ICOS beads favored the central carbon pathway through increased gluconeogenesis for bioenergetics, marked by abundant intracellular phosphoenoylpyruvate (PEP). Importantly, Th17 cells weakly stimulated with CD3/ICOS beads and redirected with a chimeric antigen receptor (CAR) that recognizes mesothelin were more effective at clearing large human mesothelioma tumors when infused into mice than those manufactured using the standard FDA-approved protocols. Taken together, these data indicate Th17 ACT therapy can be improved by using fewer activation beads during T cell manufacturing, a finding that is both cost effective and directly translatable to patients.

immunology↗