Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells
Luca Gattinoni, … , Steven A. Rosenberg, Nicholas P. Restifo
Luca Gattinoni, … , Steven A. Rosenberg, Nicholas P. Restifo
Published June 1, 2005
Citation Information: J Clin Invest. 2005;115(6):1616-1626. https://doi.org/10.1172/JCI24480.
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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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Acquisition of full effector function in vitro impairs antitumor efficac...
Acquisition of full effector function in vitro impairs antitumor efficacy. Pmel-1 CD8+ T cells at progressive stages of differentiation were generated using multiple in vitro stimulations with 1 μM hgp10025–33 and 30 IU/ml of rhIL-2. Seven days following the last stimulation, phenotypic and functional assays were performed. (A) Phenotypic differences in naive and effector subgroups. Flow cytometry analysis for the expression of lymphoid-homing molecules CD62L, CCR7, and β7 integrin; costimulatory molecule CD27; activation/effector markers CD69, CD44, CD25, and granzyme-B (GZM-B) on CD8-enriched pmel-1 naive, early effector, intermediate effector, and effector T cells. Mean fluorescence intensity (MFI) values after gating on CD8+ cells are shown. Data shown are representative of 3 independent experiments. (B) Functional differences in naive and effector subgroups. IFN-γ and IL-2 release and cytotoxic assay against hgp10025–33–pulsed MCA-205 targets. MCA-205 cells plus the irrelevant influenza nucleoprotein peptide were used as control. In the cytotoxic assay, values for control target cells (0–14%) were subtracted. Data shown are representative of 3 independent experiments. (C and D) Differentiation of antitumor T cells is inversely associated with in vivo effectiveness. WT mice bearing 10-day-old established s.c. B16 tumors were sublethally irradiated and left untreated as control or received adoptive transfer of 1 × 106 (when not otherwise indicated) CD8-enriched pmel-1 naive, early effector, intermediate effector, and effector T cells in conjunction with rFPhgp100 vaccination and exogenous rhIL-2 (36 μg per dose). Results for tumor area are the mean of measurements from 5 mice per group (± SEM). Data shown are representative of 7 independent experiments.