Immature Dendritic Cell Therapy Confers Durable Immune Modulation in an Antigen-Dependent and Antigen-Independent Manner in Nonobese Diabetic Mice

Figure 4

Spleen cell homeostatic proliferation following DC therapy. (a) Nine-week-old NOD mice received subcutaneous injection of PBS, unpulsed DCs, or ID peptide-pulsed DCs, once a week for 3 weeks. Spleen cells from NOD mice of each group at 13 weeks of age were cultured in serum-free HL-1 media alone or with DD-insulin, or ID-GAD for 86 h, and 3H-thymidine was added for incorporation during the final 16 h of culture. Proliferation was assessed by liquid scintillation quantification of counts per minute (cpm). Data shown are the mean cpm (counts per minute) of triplicate values from one of ten experiments. (b) Nine-week-old NOD mice received subcutaneous injection of PBS, unpulsed DCs, or ID peptide-pulsed DCs, once a week for 3 weeks. Spleen cells from NOD mice of each group at 40 weeks of age were cultured in serum-free HL-1 media alone or with DD-insulin, or ID-GAD for 86 h, and 3H-thymidine was added for incorporation during the final 16 h of culture. Proliferation was assessed by liquid scintillation quantification of counts per minute (cpm). Data shown are the mean cpm (counts per minute) of triplicate values from one of ten experiments. (c) Homeostatic proliferation was observed in healthy and autoimmune mouse models. NOD, B6 mice were treated with 3 weekly subcutaneous injections of DC (10⁵/injection) or PBS beginning at 9 weeks of age, and Balb/c mice were treated with intravenous injection of DC or PBS at the same age. Spleen cells were collected 2 weeks following final injection to assess 3H-thymidine proliferation in the HL-1 media in the absence of in vitro stimulation. Data shown are the mean cpm (counts per minute) ±SD.

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