Analysis of Cytokine Producing Activity of Intestinal Intraepithelial T Cells from TCR β-Chain and δ-Chain Mutant Mice

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  • Analysis of Cytokine Producing Activity
  • Analysis of Cytokine Producing Activity of Intestinal Intraepithelial T Cells from TCR β‐Chain and 5‐Chain Mutant Mice
  • Analysis of cytokine producing activity of intestinal intraepithelial T cells from TCR P-chain and β-chain mutant mice

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Abstract

Intestinal intraepithelial T cells (IELs) expressing either γδ TCR or αβ TCR have been proposed to play an important role in the regulation of intestinal epithelia by producing cytokines that directly influence the adjoining intestinal epithelial cell (IEC) functions. To illuminate this issue, we utilized TCR mutant mice to obtain γδ IELs, αβ IELs and mixed γδ and a αβ IELs from corresponding αβ T-cell-deficient (β-/-), γδ T-cell-deficient (δ-/-) and wild-type (WT) littermate mice. The production of IFN-γ by these IELs as well as the mRNA for IFN-γ, TGF-α, TGF-β1, TNF-α and TNF-β in these IELs, in conjunction with the effect of produced cytokines on the expression of class II MHC molecules by the in vitro cell line IEC-6, were investigated. IFN-γ and TGF-α specific mRNA were detectable in all freshly isolated γδ, αβ and WT IELs. In addition to the IFN-γ and TGF-α mRNA, αβ and WT IELs that had been activated in culture plates coated with anti-CD3 mAb contained mRNA for TGF-β1 and TNF-β proteins. In the cultured γδ IELs, however, the signals for IFN-γ and TGF-α transcripts were weak, and mRNA for the latter two cytokines was almost undetectable. Supernatants from in vitro culturing of αβ and WT IELs but not γδ IELs induced class II MHC gene expression in IEC-6, whereas, in the presence of anti-IFN-γ mAb, the same culture supernatants failed to do so. In fact, the concentration of IFN-γ in supernatants from αβ and WT IEL cultures was ten-to twentyfold higher than that in the supernatant from the γδ IEL culture. Finally, TGF-α specific mRNA was not detectable in the γδ and αβ IELs even after in vitro activation. These results indicate that αβ IELs are superior to γδ IELs in the ability to produce IFN-γ, TGF-β1, TGF-α and TNF-β through TCR crosslinking primary in vitro stimulation.

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