Fig. 2
The numbers of pathogenic Th17 (IL-17 producing) and IFN-γ producing cells are significantly reduced in mice treated with αDCIR2-PLP139–151 mAb compared to controls (PLP139–151 abbreviated in the figure as PLP due to space concerns). a Elispot analyses of the impact of preimmunization with αDCIR2-PLP139–151 mAb on Th17 cells. SJL/J mice were preimmunized with a low dosage (1 μg) of different fusion antibodies. Ten days later, mice were immunized with PLP139–151 and injected i.v. with pertussis toxin (200 ng) the following day to induce EAE. IL-17 ELISpot analyses were conducted on splenocytes isolated from mAb treated and untreated mice 10 days after disease induction. Splenocytes were plated onto IL-17 pre-coated plates and stimulated with 10 μg/ml PLP139–151. Wells stimulated with PHA and unstimulated wells were used as controls. Analysis was conducted using an E-biosciences IL-17 ELISpot kit. b Quantification of results of the IL-17 ELISpot assay. Pre-immunization with αDCIR2-PLP139–151 mAb (n = 2, p = 0.0059) 10 days before disease induction resulted in a decreased number of cells producing IL-17, as compared to mice that did not receive any mAbs (labeled in the figure as PLP). Number of spots per million cells was calculated by multiplying the average of triplicate wells (2 × 105) by fivefold. c IFN-γ ELISPOT analysis on splenocytes isolated from mice treated with mAb treated and untreated controls 10 days after disease induction. Splenocytes were plated onto plates pre-coated with IFN-γ and stimulated with 10 μg/ml PLP139–151. PHA and unstimulated wells were used as controls. Analysis was conducted using an IFN-γ ELISpot kit. d Quantification of results of the IFN-γ ELISpot assay. Pre-immunization with αDCIR2-PLP139–151 mAb (n = 2) resulted in a decreased number of IFN-γ producing cells, as compared to mice that had been preimmunized with ISO-PLP139–151 mAb (n = 2, p = 0.0001) or not preimmunized with either mAb (n = 2, p = 0.0001). Notably, consistent with the slight disease amelioration seen in Fig. 1, pre-treatment with ISO-PLP139–151 mAb also resulted in a reduction of IFN-γ producing cells as compared to PLP-treated mice (n = 2, p = 0.0124). Number of spots per million cells was calculated by multiplying the average of triplicate wells (2 × 105) splenocyes per well) by fivefold