IL-10–producing NKT10 cells are a distinct regulatory invariant NKT cell subset
Duygu Sag, … , Mitchell Kronenberg, Gerhard Wingender
Duygu Sag, … , Mitchell Kronenberg, Gerhard Wingender
Published July 25, 2014
Citation Information: J Clin Invest. 2014;124(9):3725-3740. https://doi.org/10.1172/JCI72308.
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Research Article Immunology

IL-10–producing NKT10 cells are a distinct regulatory invariant NKT cell subset

Abstract

Invariant natural killer T (iNKT) cells rapidly produce copious amounts of multiple cytokines after activation, thereby impacting a wide variety of different immune reactions. However, strong activation of iNKT cells with α-galactosylceramide (αGalCer) reportedly induces a hyporeactive state that resembles anergy. In contrast, we determined here that iNKT cells from mice pretreated with αGalCer retain cytotoxic activity and maintain the ability to respond to TCR-dependent as well as TCR-independent cytokine-mediated stimulation. Additionally, αGalCer-pretreated iNKT cells acquired characteristics of regulatory cells, including production and secretion of the immunomodulatory cytokine IL-10. Through the production of IL-10, αGalCer-pretreated iNKT cells impaired antitumor responses and reduced disease in experimental autoimmune encephalomyelitis, a mouse model of autoimmune disease. Furthermore, a subset of iNKT cells with a similar inhibitory phenotype and function were present in mice not exposed to αGalCer and were enriched in mouse adipose tissue and detectable in human PBMCs. These data demonstrate that IL-10–producing iNKT cells with regulatory potential (NKT10 cells) represent a distinct iNKT cell subset.

Authors

Duygu Sag, Petra Krause, Catherine C. Hedrick, Mitchell Kronenberg, Gerhard Wingender

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Figure 6

αGalCer-pretreated iNKT cells produce IL-10.

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αGalCer-pretreated iNKT cells produce IL-10. (A and B) αGalCer (1 μg) wa...
(A and B) αGalCer (1 μg) was i.v. injected into wild-type (B6) or mice i.v. injected 1 month earlier with 4 μg αGalCer (B6/αGC) (3 mice/group). Splenic iNKT cells were analyzed 90 minutes later by intracellular staining for expression of the indicated cytokines. (A) Summary graph. Data in the 2 panels are derived from 2 independent experiments. (B) Representative flow cytometric data. ICS, intracellular cytokine staining. (C and D) Control Il10GFP mice or Il10GFP mice given 4 μg αGalCer 1 month earlier (Il10GFP/αGC; αGalCer-pre) were either left untreated or were i.v. injected with 1 μg αGalCer (Stimulated), and splenic iNKT cells were analyzed 16 hours later for EGFP and CD279 (PD1) expression. Representative data (C) and a summary graph of IL-10GFP expression (D) are shown. αGalCer-pre, unstimulated = mean: 3.92% ± 0.47%; median: 3.61%. αGalCer-pre, stimulated = mean: 50.03% ± 1.51%; median: 51.71%. Control, stimulated = mean: 13.76% ± 2.04%; median: 12.85%. Data shown in D are from at least 8 independent experiments with at least 16 mice per group. Background values for IL-10 detection by intracellular staining or EGFP are shown and discussed in Supplemental Figure 8. (E) Splenic iNKT cells from the indicated mice (B6 = C57BL/6; B6/αGC = C57BL/6 mice i.v. injected 1 month earlier with 4 μg αGalCer; Il10–/– = IL-10–deficient mice) were sorted and were either left untreated or were stimulated in vitro for 4 days on αTCRβ antibody–coated plates before supernatants were collected and IL-10 levels were determined by ELISA. Representative data from 3 independent experiments are shown.