Figure 4
Unlike tumor necrosis factor (TNF)α, TNF-like cytokine 1A (TL1A) directly induces interferon (IFN)γ expression in CD3+CD56−CD161+ T cells. (a) Human peripheral blood mononuclear cells (hPBMCs) were left untreated, treated with TL1A, interleukin (IL)12, and IL18, or IL12 and IL18 plus 200 ng ml−1 TL1A for 24 h. Cells were gated on CD3+CD56 and separated into (top panel) CD161+ or (bottom panel) CD161−, and intracellular IFNγ was measured by flow cytometry. (b) hPBMCs were treated as in a, but in this case 200 ng ml−1 TNFα was also added. (c, d) hPBMCs were harvested from eight healthy volunteers and each was treated with IL12 and IL18 plus either TL1A or TNFα. (c) The percent IFNγ-positive cells and (d) the mean fluorescence intensity (MFI) of IFNγ were analyzed by flow cytometry. (e, f) hPBMCs were treated as in a, but in certain cases, anti-TL1A-0011 or anti-TNFα, plus respective isotype (iso) controls, were added to the culture. After 24 h, (e) the percentage of IFNγ CD161+ cells or (f) the MFI of IFNγ was analyzed. Each sample was run in quadruplicate. NS, not significant; NT, not receiving any treatment.
