Fig. 4

CD4+ T cells deficient in cathepsin Z exhibit proficient migration in vitro; and efficient trafficking, infiltration, and reactivation in the CNS. a To determine whether deficiency of cathepsin Z affected the ability of CD4+ T cells to undergo chemotaxis in response to CXCL9, CD4+ T cells were isolated from the spleens of WT and Cat Z^−/− mice and given 1 h to migrate through an ICAM coated Transwell plate in response to CXCL9 (n = 4). The negative control (−) well had no CXCL9. The positive control (+) well had no filter, allowing all CD4+ T cells to migrate through to the bottom of the Transwell. b To evaluate the ability of CD4+ T cells to infiltrate the CNS in WT and Cat Z^−/− mice, MOG^35–55-specific 2D2 CD4+ T cells were isolated and expanded ex vivo using IL-12 and MOG^35–55 for 48 h before adoptive transfer into WT and Cat Z^−/− recipient mice; alternatively, (c) to examine the capacity of Cat Z^−/− CD4+ T cells to infiltrate the CNS of WT mice, WT and Cat Z^−/− MOG^35–55-specific 2D2 CD4+ T cells were isolated, expanded and adoptively transferred into WT recipients. b-c Six days following adoptive transfer, the 2D2 CD4+ T cells were isolated from the CNS using a discontinuous Percoll gradient, identified by flow cytometry (CD4+, Vα3.2+) and evaluated for the expression of the activation marker CD25 (n = 4). Data presented as mean+/− SEM; no significant differences (unpaired Student’s t test, p > 0.05) from the WT control were observed