CD20 levels on young vs aged B cells, and impact of anti-CD20 treatement on T cells in EAE mice. (A) Single cells from the dura, inguinal LNs (iLNs), cervical LNs (cLNs), and spleens (Spl) of young (8 wk, n = 6) and aged (11 month, n = 6) SJL/J mice were subjected to flow cytometry analysis to assess levels of CD20 glycoprotein expression on B220^low versus B220^high B cells. Expression is represented by median fluorescence index (MFI). Data represent one experimental replicate. (B) Absolute number of CD3⁺ T cells in the femur/tibia BM, skull BM, dura, LM, and brain were analyzed by flow cytometry at day 24 of EAE (see Fig. 5) for n = 11–17 in the anti-CD20 monoclonal antibody–treated group or n = 14–20 in the isotype control antibody group. Results in B are expressed as absolute numbers of whole tissue. Results in A and B are data from two independent experiments. Error bars indicate mean ± SEM in B or SD in A. A Student’s t test (A) or Mann–Whitney test (B) was conducted to assess statistical significance (* = P < 0.05, ** = P < 0.005, *** = P < 0.0005, **** = P < 0.0005, and no label = not significant).