CD62L+ NKT cells have prolonged persistence and antitumor activity in vivo
Gengwen Tian, … , Laurence J. Cooper, Leonid S. Metelitsa
Gengwen Tian, … , Laurence J. Cooper, Leonid S. Metelitsa
Published May 16, 2016
Citation Information: J Clin Invest. 2016;126(6):2341-2355. https://doi.org/10.1172/JCI83476.
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Research Article Oncology

CD62L+ NKT cells have prolonged persistence and antitumor activity in vivo

Abstract

Vα24-invariant natural killer T cells (NKTs) localize to tumors and have inherent antitumor properties, making them attractive chimeric antigen receptor (CAR) carriers for redirected cancer immunotherapy. However, clinical application of CAR-NKTs has been impeded, as mechanisms responsible for NKT expansion and the in vivo persistence of these cells are unknown. Here, we demonstrated that antigen-induced expansion of primary NKTs in vitro associates with the accumulation of a CD62L+ subset and exhaustion of CD62L– cells. Only CD62L+ NKTs survived and proliferated in response to secondary stimulation. When transferred to immune-deficient NSG mice, CD62L+ NKTs persisted 5 times longer than CD62L– NKTs. Moreover, CD62L+ cells transduced with a CD19-specific CAR achieved sustained tumor regression in a B cell lymphoma model. Proliferating CD62L+ cells downregulated or maintained CD62L expression when activated via T cell receptor alone or in combination with costimulatory receptors. We generated HLAnull K562 cell clones that were engineered to express CD1d and costimulatory ligands. Clone B-8-2 (HLAnullCD1dmedCD86high4-1BBLmedOX40Lhigh) induced the highest rates of NKT expansion and CD62L expression. B-8-2–expanded CAR-NKTs exhibited prolonged in vivo persistence and superior therapeutic activities in models of lymphoma and neuroblastoma. Therefore, we have identified CD62L as a marker of a distinct NKT subset endowed with high proliferative potential and have developed artificial antigen-presenting cells that generate CD62L-enriched NKTs for effective cancer immunotherapy.

Authors

Gengwen Tian, Amy N. Courtney, Bipulendu Jena, Andras Heczey, Daofeng Liu, Ekaterina Marinova, Linjie Guo, Xin Xu, Hiroki Torikai, Qianxing Mo, Gianpietro Dotti, Laurence J. Cooper, Leonid S. Metelitsa

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Figure 8

aAPC-expanded NKTs have superior in vivo persistence and antitumor activity.

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aAPC-expanded NKTs have superior in vivo persistence and antitumor activ...
(A) NKTs were transduced with a CD19-specific CAR followed by expansion of transduced (CAR) or nontransduced (NT) NKTs with PBMC or B-8-2 restimulation. Four groups of mice then received i.v. injection of 2 × 105 Raji cells (day 0) followed by (day 3) i.v. injection of the respective NKT preparations (107 cells per mouse) with IL-2 (1,000 U/mouse). NKT persistence was monitored by bioluminescent imaging on the indicated days. Bioluminescent images of 7 mice per group are shown from 1 of 2 independent experiments. (B) Mice received i.v. injection of 2 × 105 luciferase-transduced Raji cells (day 0) followed by (day 2) i.v. injection of 107 NT or CAR.CD19 NKTs with IL-2 (1,000 U/mouse). (C) Mice received i.v. injection of 106 luciferase-transduced CHLA-255 cells (day 0) followed by (day 4) i.v. injection of 107 NT or CAR.GD2 NKTs with IL-2 (1,000 U/mouse). Tumor growth was monitored using bioluminescent imaging once per week. Survival plot from 2 combined experiments for the Raji model (15 mice per group) and 1 experiment with the CHLA-255 model (8 mice per group). Data were analyzed by the Kaplan-Meier method. The differences in survival were then compared using the log-rank test. ***P < 0.001.