Zhou et al. developed a two-step protocol to investigate the short-term kinetics of T cell activation. By analyzing surface markers, gene transcripts, and the single-cell secreted protein profile, they demonstrated a feedback loop in which T cell–T cell interactions following antigen stimulation led to highly polyfunctional, but not significantly differentiated T cells. Such optimally primed T cells efficiently eradicated tumors in mice, suggesting that T cell functionality and phenotype evolution can be temporally decoupled for optimization of adoptive cell therapy.
For adoptive cell transfer (ACT) immunotherapy of tumor-reactive T cells, an effective therapeutic outcome depends upon cell dose, cell expansion in vivo through a minimally differentiated phenotype, long term persistence, and strong cytolytic effector function. An incomplete understanding of the biological coupling between T cell expansion, differentiation, and response to stimulation hinders the co-optimization of these factors. We report on a biophysical investigation of how the short-term kinetics of T cell functional activation, through molecular stimulation and cell-cell interactions, competes with phenotype differentiation. T cells receive molecular stimulation for a few minutes to a few hours in bulk culture. Following this priming period, the cells are then analyzed at the transcriptional level, or isolated as single cells, with continuing molecular stimulation, within microchambers for analysis via 11-plex secreted protein assays. We resolve a rapid feedback mechanism, promoted by T cell-T cell contact interactions, which strongly amplifies T cell functional performance while yielding only minimal phenotype differentiation. When tested in mouse models of ACT, optimally primed T cells lead to complete tumor eradication. A similar kinetic process is identified in CD8+ and CD4+ T cells collected from a patient with metastatic melanoma.
Author Info: (1) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Chemistry and Chemical Engineering, Californi
Author Info: (1) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America. (2) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, United States of America. (3) Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America. (4) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America. (5) David Geffen School of Medicine, the Johnson Comprehensive Cancer Center, University of California, Los Angeles, California, United States of America. (6) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America. (7) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, United States of America. (8) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. David Geffen School of Medicine, the Johnson Comprehensive Cancer Center, University of California, Los Angeles, California, United States of America. (9) NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, United States of America. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America.
