Fig. 1.

Distinct cytokine production by NKT cell subsets. NKT cells from spleens, livers, and thymuses of B6 mice were identified as α-GC/CD1d tetramer⁺ αβTCR⁺ cells and divided further on the basis of NK1.1 and CD4 expression as shown in the upper right dot plot (shown is a representative thymus sample). Purified NKT subsets were cultured in wells coated overnight with 10 μg/ml anti-CD3 and 10 μg/ml anti-CD28. An aliquot of supernatant was harvested at 24 and 72 h to assay for secreted cytokines. IL-17 and IL-21 were assayed by ELISA. All other cytokines were assayed by cytometric bead array. The absence of bars indicates undetectable amounts of cytokine, unless “not done” is stated. For thymus and liver NKT subsets, the results are derived from 5 or 6 separate cultures collected over three independent experiments. For spleen NKT cell subsets, results are derived from 3 or 4 separate cultures collected over two independent experiments. Bars depict mean ± standard error. All cytokine values are expressed in ng/ml. Total cells = absolute cell number.