The clinical implication and molecular mechanism of preferential IL-4 production by modified glycolipid-stimulated NKT cells
Shinji Oki, … , Takashi Yamamura, Sachiko Miyake
Shinji Oki, … , Takashi Yamamura, Sachiko Miyake
Published June 1, 2004
Citation Information: J Clin Invest. 2004;113(11):1631-1640. https://doi.org/10.1172/JCI20862.
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Article Immunology

The clinical implication and molecular mechanism of preferential IL-4 production by modified glycolipid-stimulated NKT cells

Abstract

OCH, a sphingosine-truncated analog of α-galactosylceramide (αGC), is a potential therapeutic reagent for a variety of Th1-mediated autoimmune diseases through its selective induction of Th2 cytokines from natural killer T (NKT) cells. We demonstrate here that the NKT cell production of IFN-γ is more susceptible to the sphingosine length of glycolipid ligand than that of IL-4 and that the length of the sphingosine chain determines the duration of NKT cell stimulation by CD1d-associated glycolipids. Furthermore, IFN-γ production by NKT cells requires longer T cell receptor stimulation than is required for IL-4 production by NKT cells stimulated either with immobilized mAb to CD3 or with immobilized “αGC-loaded” CD1d molecules. Interestingly, transcription of IFN-γ but not that of IL-4 was sensitive to cycloheximide treatment, indicating the intrinsic involvement of de novo protein synthesis for IFN-γ production by NKT cells. Finally, we determined c-Rel was preferentially transcribed in αGC-stimulated but not in OCH-stimulated NKT cells and was essential for IFN-γ production by activated NKT cells. Given the dominant immune regulation by the remarkable cytokine production of ligand-stimulated NKT cells in vivo, in comparison with that of (antigen-specific) T cells or NK cells, the current study confirms OCH as a likely therapeutic reagent for use against Th1-mediated autoimmune diseases and provides a novel clue for the design of drugs targeting NKT cells.

Authors

Shinji Oki, Asako Chiba, Takashi Yamamura, Sachiko Miyake

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Kinetic analysis of NKT cell activation and cytokine production after gl...
Kinetic analysis of NKT cell activation and cytokine production after glycolipid stimulation. (A) Differential production of IFN-γ and IL-4 by activated NKT cells. CD3+NK1.1+ NKT cells and conventional CD3+NK1.1_ T cells were purified from liver mononuclear cells by cell sorting. The sorted cells were stimulated with immobilized mAb to CD3 for the time indicated on the x axis and were then removed and recultured on a fresh culture plate without anti-CD3 stimulation for up to 72 hours from the start of the anti-CD3 stimulation. Incorporation of [3H]thymidine (1 ∝Ci/well) during the final 16 hours of the culture was assessed (left), and culture supernatants were analyzed for the production of IL-4 (center) and IFN-γ (right) by ELISA. One experiment representative of three independent experiments with similar results is shown. (B) NKT cells purified from liver mononuclear cells were stimulated with plates coated with DimerX I loaded with αGC and were analyzed as shown in A. (C) NKT cells purified from liver mononuclear cells were stimulated as shown in A in the presence [CsA(+)] or absence [CsA(_)] of CsA (1 ∝g/ml). Culture supernatants were analyzed for the production of IL-4 and IFN-γ by ELISA.