Figure 7
From: Detection of autoreactive CD4 T cells using major histocompatibility complex class II dextramers
Ex vivo detection of MOG-reactive cells by IAb/MOG 35-55 tetramers and dextramers. (A) LNC. CD4 cells were isolated from LNC obtained from mice immunized with MOG 35-55; after treatment with NASE, cells were stained with MOG 35-55 or control tetramers or dextramers, followed by staining with anti-CD4 and 7-AAD and anti-CD25 or anti-CD44. Cells were acquired by FC, and percentages of tet+ and dext+ live (7-AAD-) cells were analyzed in CD4, CD25 and CD44 subsets. Upper and bottom left panels, respectively, represent FC plots and mean ± SEM values obtained from three experiments each involving 3 to 5 mice. (B) MNC from CNS tissues. Groups of mice were immunized with MOG 35-55 in CFA, and animals showing paralytic signs were killed and brain and spinal cords collected to harvest MNC by percoll density gradient centrifugation. Cells were stained with MOG 35-55 or control tetramers and dextramers and anti-CD4 and 7-AAD. Cells were acquired by FC and tet+ and dext+ cells were enumerated in the live CD4 subset. Representative data from three individual experiments, each involving 3 to 5 mice, are shown. Control, PLP 139-151.