Figure 3.

Flow cytometric analysis of peripheral T cells after preventive administration of GCs. (a) C57Bl/6 mice were treated three times with 100 mg/kg MP or Dex. Leukocytes were isolated from the spleens 24 hrs after the first, second, or third injection, stained for CD4 and CD3, and analyzed by flow cytometry. n = 2 (24 hrs/48 hrs), n = 5 (72 hrs). Statistical analysis for day 3 was performed using the unpaired t-test (**P < 0.01). (b) C57Bl/6 mice were treated three times at days −1, 0, and 1 with 100 mg/kg MP, 100 mg/kg Dex, or PBS as a control. At day 0, they were immunized with MOG_35-55 according to the standard protocol. 12 days after immunization, leukocytes were isolated from the spleens, and absolute T-cell numbers were calculated based on flow cytometric analysis after staining for CD4 and CD8. n = 6. Statistical analysis was performed using the unpaired t-test (**P < 0.01). (c) The relative expression level of LFA-1 was determined after three injections of MP or Dex in splenic T cells using the same mice as in (A). The mean fluorescence intensity (MFI) 24 hrs after the last injection is depicted; n = 5 for each group. All values are depicted as mean ± SEM. Statistical analysis was performed using the unpaired t-test (**P < 0.01). (d) The percentage of FoxP3⁺ CD4⁺ CD25⁺ splenic T cells amongst all CD4⁺ T cells was determined 24 hrs after the last injection by flow cytometry; n = 5 for each group. All values are depicted as mean ± SEM. Statistical analysis was performed using the unpaired t-test.