Journal: Nature Communications
Article Title: Altered thymic differentiation and modulation of arthritis by invariant NKT cells expressing mutant ZAP70
Figure Lengend Snippet: NKT1 cells negatively correlate with arthritis severity in SKG mice. a Representative flow cytometry plot of cytokine profile of iNKT (TCRβ int CD1d-αGC tetramer + ) cells from joint-draining lymph nodes and joints (wrists and ankles) of arthritic SKG mice stimulated with PMA and ionomycin. b Representative flow cytometry of iNKT (TCRβ int CD1d-αGC tetramer + ) cell subsets in ankles during the course of mannan-induced arthritis in male SKG mice. Day 0 (circle), day 12 (square), day 22 (triangle), and day 47 (diamond) post-mannan injection. c Frequency of NKT1 (T-bet + RORγt − , left, green) and NKT17 cells (T-bet − RORγt + , right, purple) during the course of mannan-induced arthritis in SKG mice. d Correlation between NKT1 (left panel, green) and NKT17 (right panel, purple) and changes in ankle swelling in mice with mannan-induced arthritis. e Representative flow cytometry plot of iNKT cells (CD3 + Vα24Jα18 + (6B11)) in paired peripheral blood (PB) and synovial fluid (SF) samples derived from rheumatoid arthritis (RA) patients ( n = 8). f Levels of IFNγ in paired peripheral blood (PB) and synovial fluid (SF) samples stimulated with αGC (RA patients, n = 4). g Representative histogram (of a total of 5 RA patients) of IFN-γ expressing iNKT cells (CD3 + Vα24Jα18 + (6B11 antibody-reactive) after stimulation with PMA and ionomycin (black/red histogram) or unstimulated control (filled histogram). h Expression of IFN-γ and IL-4 from splenic iNKT cells after injection of αGC in WT BALB/c (blue) or SKG mice (red). i Expression of IFN-γ by splenic iNKT cells after injection of LPS in WT BALB/c (blue) or SKG mice (red). j Serum IFNγ in WT BALB/c (blue) or SKG mice (red) after LPS injection. k Production of IFN-γ by iNKT cells after stimulation of lymph node cultures for 48 h with IL-12 (20 ng/ml). Lymph node cultures prepared from WT BALB/c (blue), pre-arthritic SKG (red, 7–8 weeks old) and arthritic SKG mice (black, 40 days post-mannan injection). Representative of at least two independent experiments are shown. Graphs represent mean ± SEM with symbols representing individual mice. * p
Article Snippet: For evaluation of IFN-γ production by iNKT cells directly ex vivo, freshly isolated PBMC and SFMC were incubated for 4 h with phorbol 12-myristate 13-acetate (PMA, 25 ng/ml), calcium-ionomycin (CaI, 1 µg/ml; both from Sigma) and brefeldin A (BFA, 10 ng/ml; BD) or BFA alone (negative control).
Techniques: Mouse Assay, Flow Cytometry, Cytometry, Injection, Derivative Assay, Expressing