Nonclassical CD1d-restricted NK T cells that produce IL-13 characterize an atypical Th2 response in ulcerative colitis
J. Clin. Invest. Ivan J. Fuss, et al. 113:1490 doi:10.1172/JCI19836 [Go to this article.]

Figure 2
Intracellular cytokine expression of CD161⁺ LPMC. (A) CD161 expression on unstimulated CD3⁺ LPMCs. LPMCs from control (n = 5), involved CD (n = 6), and involved UC (n = 8) tissues were stained with fluorochrome-labeled Ab’s recognizing various surface antigens (CyChrome-labeled anti-CD3, PE-labeled anti-CD4, and FITC-labeled anti-CD161) and then were analyzed for the expression of CD4⁺ versus CD161⁺ cells on CD3-gated cells. (B) Intracellular IL-13 cytokine production by stimulated LPMCs. LPMCs from control (n = 5), involved CD (n = 6), and involved UC (n = 8) tissues were cultured for 48 hours with anti-CD2/anti-CD28 with addition of monensin for the last 6 hours of culture. The resultant cells were stained with fluorochrome-labeled Ab’s recognizing various surface antigens (CyChrome-labeled anti-CD3, APC-labeled anti-CD8, and FITC-labeled anti-CD161) and then were fixed/permeabilized prior to intracellular cytokine staining with PE-labeled anti_IL-13. Cells were gated on CD3 and analyzed for the expression of CD8^_ (CD4⁺) cells and CD161⁺ cells versus IL-13⁺ cells. (C) Intracellular IFN-γ cytokine production by stimulated LPMCs was the same as in B, except LPMCs were cultured for 6 hours with PMA and ionomycin with the addition of monensin for the last 5 hours of culture, and fixed/permeabilized cells were stained with APC-labeled anti_IFN-γ. (D) Intracellular IL-13 cytokine production by unstimulated LPMCs. LPMCs from involved CD (n = 4) and involved UC (n = 4) tissues were stained with fluorochrome-labeled Ab’s recognizing various surface antigens (CyChrome-labeled anti-CD3, APC-labeled anti-CD8, and FITC-labeled anti-CD161) and then fixed/permeabilized prior to intracellular cytokine staining with PE-labeled anti_IL-13. Cells were gated on CD3 and analyzed for the expression of CD4⁺ cells and CD161⁺ cells versus IL-13⁺ cells.