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. 2009 Sep 25;5(9):e1000588. doi: 10.1371/journal.ppat.1000588

Figure 4. LT inhibits α-GC-stimulated cytokine production.

Figure 4

A C57BL/6 mice were treated with 100 µg of LT in PBS by the i.v. route or mock-treated with PBS alone. After 4 d, splenocytes were obtained and stimulated in vitro with α-GC at a final concentration of 50 ng/ml. Supernatants were collected after a further 24 and 48 h and stored at −80°C. B Splenocytes from Jα18−/− mice and ex vivo-expanded NKT cells from C57BL/6 mice were treated in vitro with LT at a final concentration of 1 µg/ml for 1 h before washing and culturing separately or together in the presence or absence of α-GC as described in A. IL-4 and IFNγ concentrations in the supernatants were then determined by Bio-Plex analysis. Data show mean cytokine concentration for 3 mice per group ±SD. Asterisk indicates significant difference between cytokine concentration in un-treated control and samples from LT-treated mice. C C57BL/6 mice were treated with PBS, non-functional LT mutant, or wild type LT before enrichment of NKT cells using anti-NK1.1-based magnetic isolation. Cells were stimulated with anti-CD3 and CD28 mAbs and culture supernatants collected after 48 h.