Cytokine therapy reverses NK cell anergy in MHC-deficient tumors
Michele Ardolino, … , K. Christopher Garcia, David H. Raulet
Michele Ardolino, … , K. Christopher Garcia, David H. Raulet
Published October 20, 2014
Citation Information: J Clin Invest. 2014;124(11):4781-4794. https://doi.org/10.1172/JCI74337.
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Research Article

Cytokine therapy reverses NK cell anergy in MHC-deficient tumors

Abstract

Various cytokines have been evaluated as potential anticancer drugs; however, most cytokine trials have shown relatively low efficacy. Here, we found that treatments with IL-12 and IL-18 or with a mutant form of IL-2 (the “superkine” called H9) provided substantial therapeutic benefit for mice specifically bearing MHC class I–deficient tumors, but these treatments were ineffective for mice with matched MHC class I+ tumors. Cytokine efficacy was linked to the reversal of the anergic state of NK cells that specifically occurred in MHC class I–deficient tumors, but not MHC class I+ tumors. NK cell anergy was accompanied by impaired early signal transduction and was locally imparted by the presence of MHC class I–deficient tumor cells, even when such cells were a minor population in a tumor mixture. These results demonstrate that MHC class I–deficient tumor cells can escape from the immune response by functionally inactivating NK cells, and suggest cytokine-based immunotherapy as a potential strategy for MHC class I–deficient tumors. These results suggest that such cytokine therapies would be optimized by stratification of patients. Moreover, our results suggest that such treatments may be highly beneficial in the context of therapies to enhance NK cell functions in cancer patients.

Authors

Michele Ardolino, Camillia S. Azimi, Alexandre Iannello, Troy N. Trevino, Lucas Horan, Lily Zhang, Weiwen Deng, Aaron M. Ring, Suzanne Fischer, K. Christopher Garcia, David H. Raulet

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Figure 3

NK cells infiltrating MHC class I–deficient tumors have impaired functional responsiveness.

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NK cells infiltrating MHC class I–deficient tumors have impaired functio...
(A–C) Fourteen days after implantation, the percentages of NK cells among the viable cells in the tumors (A) and their responsiveness (B and C) were assessed by flow cytometry. Infiltrating leukocytes were restimulated in vitro with NKR-P1C antibody or control IgG (indicated as “+” and “–” in the legends), and IFN-γ accumulation and/or CD107a expression were determined. (D) Functional responsiveness of NK cells infiltrating tumors derived from RMA, RMA-S, or RMA-S/Tap2 cells was assayed as described in A. (E) RMA or RMA-S cells were injected in a Matrigel solution 7, 10, or 14 days before the functional assays were performed. (F) Tumor-infiltrating leukocytes were restimulated in vitro with NKp46 antibody or control IgG, and IFN-γ accumulation and CD107a expression were evaluated. (G) RMA or RMA-S cells were implanted in Ubi-GFP/BL6 hosts, and after 14 days tumor-infiltrating leukocytes were sorted and cocultured for 5 hours with YAC-1 cells at an effector/target ratio of 10:1. NK cell degranulation was determined by assaying of CD107a expression at the cell surface. (H) Leukocytes from RMA or RMA-S tumors were stimulated with PMA and ionomycin before IFN-γ accumulation on NK cells was measured by flow cytometry. In all graphs, bars represent means ± SD. In all panels NK cells were gated as viable-CD3–CD19–Ter119–NKp46+ cells. The experiments included at least 4 mice per group and were performed 10 (B), 3 (D and F), or 2 (E, G, and H) times with similar results. Statistical analyses were performed with the 2-tailed unpaired Student’s t test.