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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Research Article Immunology

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 3

Heat-killed E. coli induces hyporesponsiveness of iNKT cells to α-GalCer rechallenge ex vivo.

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Heat-killed E. coli induces hyporesponsiveness of iNKT cells to α-GalCer...
(A) In vivo dynamics of the iNKT cell response. Mice were injected with α-GalCer (5 μg/mouse, i.p.) or heat-killed E. coli and sacrificed at the indicated time points, and spleen and liver mononuclear cells were prepared and stained for the identification of iNKT cells. (B) Total spleen iNKT cell counts and percentage of liver iNKT cells at the indicated times, for a total of 5–10 mice per group, pooled from 2 separate experiments. *P < 0.05 compared with naive animals. (C) α-GalCer recall response of mice at the indicated times after treatment with heat-killed E. coli. Mice were injected with α-GalCer or heat-killed E. coli and sacrificed 3, 4, or 6 weeks later, and splenocytes 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. *P < 0.05 compared with naive splenocytes. (D) Proliferative defect in iNKT cells from mice treated with heat-killed E. coli. Spleen cells from naive mice or from mice injected 4 or 6 weeks earlier with α-GalCer or heat-killed E. coli were labeled with CFSE. Cells were then cultured with α-GalCer (100 ng/ml) for 24 hours, then washed and cultured for an additional 96 hours without α-GalCer. At the end of the culture period, cells were harvested, stained, and analyzed by flow cytometry. (E) iNKT cell cytokine production. Spleen cells were prepared at the indicated time points, and cells were cultured for 6 hours in vehicle or 100 ng/ml α-GalCer in the presence of GolgiPlug. Cells were then harvested and stained.