Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells
Luca Gattinoni, Christopher A. Klebanoff, Douglas C. Palmer, Claudia Wrzesinski, Keith Kerstann, Zhiya Yu, Steven E. Finkelstein, Marc R. Theoret, Steven A. Rosenberg, Nicholas P. Restifo
Luca Gattinoni, Christopher A. Klebanoff, Douglas C. Palmer, Claudia Wrzesinski, Keith Kerstann, Zhiya Yu, Steven E. Finkelstein, Marc R. Theoret, Steven A. Rosenberg, Nicholas P. Restifo
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Research Article Immunology

Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells

Abstract

T cell differentiation is a progressive process characterized by phenotypic and functional changes. By transferring tumor-specific CD8+ T cells into tumor-bearing mice at various stages of differentiation, we evaluated their efficacy for adoptive immunotherapy. We found that administration of naive and early effector T cells, in combination with active immunization and IL-2, resulted in the eradication of large, established tumors. Despite enhanced in vitro antitumor properties, more-differentiated effector T cells were less effective for in vivo tumor treatment. Several events may underlie this paradoxical phenomenon: (a) downregulation of lymphoid-homing and costimulatory molecules; (b) inability to produce IL-2 and access homeostatic cytokines; and (c) entry into a proapoptotic and replicative senescent state. While the progressive acquisition of terminal effector properties is characterized by pronounced in vitro tumor killing, in vivo T cell activation, proliferation, and survival are progressively impaired. These findings suggest that the current methodology for selecting T cells for transfer is inadequate and provide new criteria for the generation and the screening of optimal lymphocyte populations for adoptive immunotherapy.

Authors

Luca Gattinoni, Christopher A. Klebanoff, Douglas C. Palmer, Claudia Wrzesinski, Keith Kerstann, Zhiya Yu, Steven E. Finkelstein, Marc R. Theoret, Steven A. Rosenberg, Nicholas P. Restifo

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Subfractionated CD62Lhigh and CD62Llow early effector cells possess simi...
Subfractionated CD62Lhigh and CD62Llow early effector cells possess similar properties in vitro. (A and B) Phenotypic characterization of CD62Lhigh and CD62Llow subsets. Five days after the initial stimulation with antigen and IL-2, CD62Lhigh and CD62Llow populations were sorted by MACS CD62L-positive selection column and analyzed by flow cytometry on day 6 for the expression of IL-7Rα; lymphoid-homing molecules β7 integrin and CCR7; the costimulatory molecule CD27; activation/effector markers CD44, CD69, CD25, and granzyme-B; and the senescence marker KLRG-1. MFI values after gating on CD8+Vβ13+ cells are shown. The MFI was determined for all groups at the same time under the same cytoflourometric settings using the same reagents and isotype-matched controls. Data shown are representative of 2 independent experiments. (C) CD62Lhigh and CD62Llow subsets have similar functional qualities. 51Cr, IFN-γ, and IL-2 release against hgp10025–33–pulsed MCA-205 targets. MCA-205 cells plus influenza nucleoprotein peptide (np) were used as control. Where error bars are not visible, it is because they are obscured by the symbols used. Data shown are representative of 2 independent experiments. (D) CD62Lhigh and CD62Llow subsets have similar proliferative capacity. CD62Lhigh and CD62Llow cells were labeled with CFSE and were cultured with hgp10025–33 peptide–pulsed on irradiated splenocytes in the presence of 30 IU/ml of rhIL-2. Cells were harvested from day 1 to day 4, and their CFSE content was determined. Results after gating on CD8+Vβ13+ population are shown. This experiment was performed twice, with similar results.