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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: J Autoimmun. 2011 Apr 12;37(1):28–38. doi: 10.1016/j.jaut.2011.03.001

Figure 1. Regulatory phenotype of NKT-instructed APCs.

Figure 1

A) NKT-instructed APCs limit the proliferation of allogeneic T cells. Purified peripheral blood T cells were labeled with CFSE and cultured for 7 days with allogeneic NKT-instructed or cytokine-induced APCs. The plot shows percentages of live T cells that underwent cell division as estimated by dilution of CFSE signal; each pair of symbols represents an independent analysis of an allogeneic T cell-APC mixture. B) Cytokine-induced and NKT-instructed APCs produce similar levels of arginase. APCs were stimulated with LPS for 24 hours or left unstimulated, and the culture supernatants were assessed for arginase activity using a chromogenic assay that detects urea generated by the presence of enzyme (QuantiChrom, Bioassay Systems Inc.). C and D) APCs were matured with LPS for 8 hours, washed and cultured with allogenic T cells in the presence of the indicated concentrations of either the arginase inhibitor Nor-NOHA (B), or the iNOS inhibitor 1400W (C), for 7 days. Proliferation is shown as the percent of the live T cells (DAPI-/CD3+) that showed reduced CFSE intensity. Results shown are from one representative experiment out of 2 independent analyses. E) NKT-instructed APCs do not induce increased numbers of regulatory T cells. Flow cytometric analysis of intracellular FoxP3 expression by allogeneic T cells after 7 days of culture with cytokine-induced or NKT-instructed APCs. Similar results were obtained in 3 independent experiments.