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. 2012 Feb 7;10(2):e1001255. doi: 10.1371/journal.pbio.1001255

Figure 3. Differential gene expression and cytokine production among thymic iNKT cell subtypes from B6 mice.

Figure 3

(A, B, D, H) Quantitative PCR analysis of thymic iNKT subtypes. Thymic iNKT cells further divided into four subtypes based on the expression of CD4 and IL-17RB (red, CD4− IL-17RB+; orange, CD4+ IL-17RB+; blue, CD4− IL-17RB−; green, CD4+ IL-17RB−). One representative out of three experiments is shown (mean ± SEM). (A) The purity of the sorted cells was confirmed by the relative Il17rb and Cd4 mRNA expression levels in the respective subtypes. Il2rb ( = Cd122) expression was restricted to CD4− and CD4+, IL-17RB− iNKT cells. (B) Expression of TH1/TH2/TH17 related genes. TH1 related: Ifng, Tbx21 and Stat4, TH2 related; Il4 and Gata3, and TH17 related: Il17a, Il22 and Rorc transcripts were analyzed. (D) Expression of cytokine receptor genes. Receptor for IL-12, IL-23, and IL-25 were analyzed. The component chains of the various receptors are IL-12 receptor: IL-12Rβ2/IL-12Rβ1; IL-23 receptor: IL-23R/IL-12Rβ1; IL-25 receptor: IL-17RB/IL-17RA. (H) Expression of chemokine receptor genes. Ccr4, Ccr6, Ccr7, Cxcr3, and Cxcr6. (C, E, F, G) In vitro cytokine production by thymic iNKT cell subtypes (red, CD4− IL-17RB+; orange, CD4+ IL-17RB+; blue, CD4− IL-17RB−; green, CD4+ IL-17RB−). Sorted thymic iNKT subtypes (5×104 cells/100 µL) were co-cultured with BM-DCs (5×103/100 µL) for 48 h in the presence of α-GalCer (100 ng/µL) (C), IL-12 (10 ng/µL) (E), IL-23 (10 ng/µL) (F), or IL-25 (10 ng/µL) (G). Levels of IFN-γ, IL-4, IL-9, IL-10, IL-13, IL-17A, and IL-22 were analyzed. The data are representative of three independent experiments (mean ± SEM).