PMC:3279418 / 16693-19345 JSONTXT

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The reduced Th1 differentiation following Pam3CYS stimulation is consistent with the recent report of decreased IL12 production in cpdm macrophages following TLR2 stimulation [31]. In contrast, more Th2-specific IL4 cytokine was produced in the cpdm BMDC co-cultures than the WT control (Fig. 7B), suggesting Th2-skewed immunogenicity of cpdm BMDC. The production of Th17-specific cytokine IL17A following LPS stimulation of dendritic cells was similar between stimulated WT and cpdm BMDC cocultures (data not shown). Despite the distinct Th1- and Th2-stimulating abilities of WT and cpdm BMDC, they were equally effective in IL2 production from BMDC-T cell co-cultures except for poly I:C stimulation where WT BMDC induced more IL2 than cpdm BMDC (Fig. 7C). The Th2-biased stimulating capability of cpdm BMDC when co-cultured with allogeneic naïve CD4+ T cells is consistent with the Th2-dominant cytokine phenotype observed in cpdm mice.\n10.1371/journal.pone.0031809.g007 Figure 7 Stimulated cpdm BMDC induced Th2-biased cytokine production from naïve CD4+ T cells.\nWT and cpdm BMDC (5×104 cells in 0.1 mL complete medium; MHC haplotype: H-2b) were incubated with medium, 100 ng/mL LPS, 25 µg/mL poly I:C or 5 µg/mL Pam3CYS. After 24 hours, cells were washed with PBS and incubated with freshly isolated allogeneic naïve CD4+ T cells (2.5×105 cells in 0.1 mL complete medium; MHC haplotype: H-2d). After 5 days, supernatants were collected and the secretion of IFNγ, IL4, and IL2 were measured by ELISA. Negative controls are 1) stimulated BMDC without co-culture with allogeneic CD4+ T cells; 2) allogeneic CD4+ T cells without co-culture with stimulated BMDC. Samples from both negative controls had no detectable production of the aforementioned cytokines (not shown). 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