Figure 6.

Passive EAE is less severe in Klrk1^−/− than in wild type recipients. Leukocytes from MOG-immunized C57BL/6 were reactivated in vitro and then adoptively transferred to Klrk1^−/− (black) and wild type (Klrk1^+/+; white) recipients. (A) Mice were followed for clinical score. Mean ± SEM, n = 9–10. 2way ANOVA Klrk1^−/− vs. Klrk1^+/+ day 22–23 ^*p < 0.05 for all other days from day 12 to the end: ^***p < 0.001. (B) Flow cytometry analysis of ex vivo expanded T lymphocytes from MOG-immunized donor mice. Lymph node cells were collected, labeled with CFSE, and put in culture as described in materials and methods in the absence or presence of MOG₃₅₋₅₅ for 72 h. PMA, ionomycin, brefeldin A and monensin were added for 4 h prior to flow cytometry staining and analysis for intracellular mediators: GM-CSF, IFNγ, IL-17 and Granzyme B (GrB) as indicated. Contour plots illustrate gated events on CD4 or CD8 T cells expanded in vitro in the absence (Nil) or presence of MOG. (C) Representative detection of demyelination using fluoromyelin and DAPI in spinal cord sections from Klrk1^−/− and Klrk1^+/+ mice. White arrows indicate zones of myelin loss. (D) Western blot detection of MULT1 and albumin in CSF from Klrk1^−/− and Klrk1^+/+ mice 50 days after the adoptive transfer of activated lymphocytes. One representative western blot is illustrated and quantification of 55 kDa and 25 kDa forms as relative expression compared to albumin Mean ± SEM n = 7. Statistical differences for relative abundance of MULT1 25kDa form Klrk1^+/+ vs. Klrk1^{−/− **}p < 0.01.