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. Author manuscript; available in PMC: 2015 May 1.
Published in final edited form as: J Autoimmun. 2014 Feb 7;50:87–98. doi: 10.1016/j.jaut.2014.01.002

Figure 2.

Figure 2

BW iNKT cells, produce significantly higher amounts of IL-4 and IL-17 than Tcon cells and Ly108lo iNKT cells are predominant IL-17 producing iNKT cells. Cells were sorted from three pooled spleens of young mice and cultured (5X105/ml) for 3 days with or without plate bound anti-CD3 and CD28 mAbs, culture supernatants were then collected for IFNγ, IL-4 and IL-17 measurement by ELISA or Luminex. (A) IFNγ, IL-4 and IL-17 concentrations in supernatants of stimulated iNKT and Tcon cells from BW mice. Cytokines measurements are shown on the right. (B) IFNγ, IL-4 and IL-17 production by BW CD4+ Ly108hi, CD4+ Ly108lo and CD4− Ly108lo iNKT cells. Sorting gates are shown on the left, and cytokines measurements were shown on the right. (C) IFNγ, IL-4 and IL-17 production by BW NK1.1+ and NK1.1− iNKT cells. FACS profile is shown for the sorting gates on the left, and cytokines measurements are shown on the right. (D) IFNγ, IL-4 and IL-17 production by BW NK1.1−Ly108hi and NK1.1−Ly108lo iNKT cells. FACS profile is shown for the sort gates on the left, and cytokines measurements are shown on the right. Group differences with P>0.05 was not considered statistically significant (ns). *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001 (two-tailed Student’s t-test). Data (mean ± s.e.m.) are calculated from four independent experiments (A, B) or derived from triplicate cultures collected from one representative experiment of three with similar results (C, D). Cytokine concentrations without anti-CD3 and CD28 mAbs were less than 100pg/ml.