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Small intestinal CD8+TCRγδ+NKG2A+ intraepithelial lymphocytes have attributes of regulatory cells in patients with celiac disease
Govind Bhagat, … , Peter H.R. Green, John S. Manavalan
Govind Bhagat, … , Peter H.R. Green, John S. Manavalan
Published December 6, 2007
Citation Information: J Clin Invest. 2008;118(1):281-293. https://doi.org/10.1172/JCI30989.
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Research Article Immunology

Small intestinal CD8+TCRγδ+NKG2A+ intraepithelial lymphocytes have attributes of regulatory cells in patients with celiac disease

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Abstract

Intraepithelial lymphocytes (IELs) bearing the γδ TCR are more abundant in the small intestinal mucosa of patients with celiac disease (CD) compared with healthy individuals. However, their role in disease pathogenesis is not well understood. Here, we investigated the functional attributes of TCRγδ+ IELs isolated from intestinal biopsies of patients with either active celiac disease (ACD) or those on a gluten-free diet (GFD). We found that compared with individuals with ACD, individuals on GFD have a higher frequency of CD8+TCRγδ+ IELs that express the inhibitory NK receptor NKG2A and intracellular TGF-β1. TCR triggering as well as cross-linking of NKG2A increased both TGF-β1 intracellular expression and secretion in vitro. Coculture of sorted TCRγδ+NKG2A+ IELs, IL-15–stimulated TCRαβ+ IELs, and HLA-E+ enterocytes resulted in a decreased percentage of cytotoxic CD8+TCRαβ+ IELs expressing intracellular IFN-γ and granzyme-B and surface NKG2D. This inhibition was partially abrogated by blocking either TGF-β alone or both NKG2A and HLA-E. Thus, our data indicate that suppression was at least partially mediated by TGF-β secretion as a result of engagement of NKG2A with its ligand, HLA-E, on enterocytes and/or TCRαβ+ IELs. These findings demonstrate that human small intestinal CD8+TCRγδ+ IELs may have regulatory potential in celiac disease.

Authors

Govind Bhagat, Afzal J. Naiyer, Jayesh G. Shah, Jason Harper, Bana Jabri, Timothy C. Wang, Peter H.R. Green, John S. Manavalan

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Figure 5

Frequency of CD3+CD8+TCRγδ+ IELs expressing intracellular TGF-β1 increases in patients on GFD.

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Frequency of CD3+CD8+TCRγδ+ IELs expressing intracellular TGF-β1 increas...
(A) Representative flow cytometry histograms showing CD3+TCRαβ+ IELs and CD3+TCRγδ+ IELs expressing intracellular IL-10 and TGF-β1 and cell-surface LAP TGF-β1 in jejunal biopsies from a GFD patient. Cells were stimulated for 4 hours with PMA and ionomycin in the presence of brefeldin A. (B) Box-and-whisker plots comparing percentages of CD3+TCRαβ+ IELs, CD3+CD8+TCRγδ+ IELs, and CD3+CD8–TCRγδ+ IELs expressing intracellular TGF-β1, from ACD patients (n = 15), GFD patients (n = 23), and controls (n = 10). (C) Box-and-whisker plots comparing percentages of CD3+TCRαβ+ IELs, CD3+CD8+TCRγδ+ IELs, and CD3+CD8–TCRγδ+ IELs expressing cell-surface LAP TGF-β1, from ACD patients (n = 8), GFD patients (n = 11), and controls (n = 7). (D) Representative flow cytometry histograms showing percentage of CD3+TCRγδ+ NKG2A+ IELs and CD3+TCRγδ+ NKG2A– IELs expressing intracellular TGF-β1 and cell-surface LAP TGF-β1 in jejunal biopsies from a GFD patient. (E) Representative flow cytometry histograms showing percentage of CD3+TCRγδ+ IELs (from an individual on GFD) expressing intracellular TGF-β1 (top row) and cell-surface LAP TGF-β1 (bottom row) after 24 hours culture in wells coated with 10 μg/ml mouse IgG1, 50 μg/ml anti-human NKG2A (clone 131411), or 50 μg/ml anti-human NKG2A plus 10 μg/ml anti-human CD3 (UCHT1; data representative of 3 independent experiments). (F) Levels (mean ± SEM pg/ml) of TGF-β1 in supernatants taken from cultures of CD3+TCRγδ+ IELs that were incubated for 24 hours in wells coated with 10 μg/ml mouse IgG1, 50 μg/ml anti-human NKG2A (clone 131411), or 50 μg/ml of anti-human NKG2C (clone 134522). (G) Levels (mean ± SEM pg/ml) of TGF-β1 in supernatants taken after 48 hours of cocultures of CD3+TCRγδ+ NKG2A+ IELs with HLA-E+CD3+TCRαβ IELs, CD3+TCRγδ+NKG2A– IELs with HLA-E+CD3+TCRαβ IELs, or CD3+TCRγδ+NKG2A+ IELs with HLA-E– CD3+TCRαβ IELs.

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