NKT Cells Stimulated by Long Fatty Acyl Chain Sulfatides Significantly Reduces the Incidence of Type 1 Diabetes in Nonobese Diabetic Mice
Figure 3
Analyses of NOD T cell proliferative responses stimulated by sulfatides.
(A) Ability of C24:0 sulfatide to protect from T1D does not correlate with its capacity to stimulate a CD4+ T cell proliferative response. Spleen CD4+ T cells from female NOD mice (3–5 week-old) were co-cultured for 72 h in vitro in the presence of mitomycin C treated CD4– cells at CD4+:CD4– ratios of 1∶1, 10∶1 or 100∶1 with either control vehicle, aGalCer (100 ng/ml) or varying concentrations (5–50 mg/ml) of sulfatide (C16:0 or C24:0). [3H]-thymidine (1 mCi/well) was added for the final 18 h of culture before harvesting the cells and quantitating their thymidine incorporation. Data shown were obtained from one of two representative and reproducible experiments. (B) Proliferative responses of splenocytes from non-diabetic and diabetic NOD mice (2–4 mice/group) in response to in vitro stimulation by sulfatide or aGalCer. Splenocytes (8×105) were incubated in a 96-well plate with a titrated concentration of sulfatide or aGalCer, and [3H]-thymidine incorporation in triplicate wells was determined following culture for 90 h, as previously described [2]. No detectable response was found to another control glycolipid mGM1 in these assays. Data from one of three representative experiments are shown.