Impact of bacteria on the phenotype, functions, and therapeutic activities of invariant NKT cells in mice
Sungjune Kim, … , Lan Wu, Luc Van Kaer
Sungjune Kim, … , Lan Wu, Luc Van Kaer
Published May 1, 2008
Citation Information: J Clin Invest. 2008;118(6):2301-2315. https://doi.org/10.1172/JCI33071.
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Impact of bacteria on the phenotype, functions, and therapeutic activities of invariant NKT cells in mice

Abstract

Invariant NKT (iNKT) cells are innate-like lymphocytes that recognize glycolipid antigens in the context of the MHC class I–like antigen-presenting molecule CD1d. In vivo activation of mouse iNKT cells with the glycolipid α-galactosylceramide (α-GalCer) results in the acquisition of a hyporesponsive (anergic) phenotype by these cells. Because iNKT cells can become activated in the context of infectious agents, here we evaluated whether iNKT cell activation by microorganisms can influence subsequent responses of these cells to glycolipid antigen stimulation. We found that mouse iNKT cells activated in vivo by multiple bacterial microorganisms, or by bacterial LPS or flagellin, became unresponsive to subsequent activation with α-GalCer. This hyporesponsive phenotype of iNKT cells required IL-12 expression and was associated with changes in the surface phenotype of these cells, reduced severity of concanavalin A–induced hepatitis, and alterations in the therapeutic activities of α-GalCer. These findings may have important implications for the development of iNKT cell–based therapies.

Authors

Sungjune Kim, Saif Lalani, Vrajesh V. Parekh, Tiffaney L. Vincent, Lan Wu, Luc Van Kaer

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Figure 6

Role of bacterial TLR ligands and IL-12 in the induction of iNKT cell hyporesponsiveness.

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Role of bacterial TLR ligands and IL-12 in the induction of iNKT cell hy...
(A) Mice were injected with E. coli flagellin (20 μg) or LPS (3 doses of 10 μg every 3 days) and 3 weeks later rechallenged in vivo with vehicle (veh) or α-GalCer (1 μg/mouse, i.p.). Mice were sacrificed 3 days later, and spleen cells were stained. (B) Total spleen iNKT cells calculated from the experiments shown in A. (C) In vitro response of splenocytes to α-GalCer from mice treated 3 weeks earlier with α-GalCer, E. coli flagellin, or E. coli LPS. Mice were injected with α-GalCer or the indicated TLR ligands and sacrificed 3 weeks later, and splenocytes (2 × 105 per well) were cultured with graded doses of α-GalCer. After 3 days, proliferation was assessed, and culture supernatants were evaluated for IL-4 and IFN-γ. (D) Mice were injected with Salmonella flagellin and LPS (3 doses of 10 μg every 3 days) and 3 weeks later rechallenged in vivo with vehicle or α-GalCer (1 μg/mouse, i.p.). Mice were sacrificed 3 days later, and spleen cells were stained. (E) Total spleen iNKT cells calculated from the experiments shown in D. (F and G) E. coli–induced iNKT cell hyporesponsiveness is dependent on IL-12. (F) Wild-type and IL-12p40–deficient mice were treated with α-GalCer or heat-killed E. coli. Three weeks later, mice were sacrificed and splenocytes cultured with graded doses of α-GalCer; proliferation was assessed, and culture supernatants were evaluated for IL-4 and IFN-γ. (G) Spleen cells were prepared at the indicated time points and cultured for 6 hours with vehicle or 100 ng/ml α-GalCer in the presence of GolgiPlug. Cells were then harvested and stained. Data in B, C, E, and F are presented as mean ± SEM. *P < 0.05 compared with naive mice.