Figure 5: Proliferating CD4+, CD8+ and B220+ cells upon TT stimulation from BALB/c mice treated or untreated with RA (20 μg/dose/day) or its vehicle (sesame oil), intranasally immunized with TT (1 μg/dose) and sistemically (i.p.) boosted with TT and Alum. After 8 months cells collected from spleens (A) and mesenteric lymph nodes (B) were cultured in vitro in the presence or in the absence of TT (1μg/ml) or anti-CD3 (αCD3) mAb for 5 days and the proliferating cells were detected by CFSE assay. Cells were labelled with CFSE, cultured in vitro as described, stained with fluorocrome conjugated anti-CD4, anti-CD8 or anti-B220 mAb and analyzed by flow cytometry.