Figure 3. LIGHT is required for lymphocytes and DC migration to inflamed LN.

WT and LIGHT^−/− mice were immunized with CFA. A. Immediately after CFA immunization, Ly5.1 WT splenocytes were adoptively transferred to mice. Accumulation of Ly5.1+ cells in iLN was analyzed 24 hours after immunization. *, P < 0.05, **, P < 0.01. Representative of two experiments. B. Ly5.1 splenocytes were treated with FTY720 0.5μg/ml for 1hr before adoptive transfer and CFA immunization in WT versus LIGHT^−/− mice. 24 hours after CFA immunization, accumulation of Ly5.1 T and B cells in DLN were counted by flow cytometry. *, P < 0.05, ***, P < 0.001. Representative of two experiments. C. Immediately after CFA immunization, Ly5.1 WT splenocytes were adoptively transferred i.v. to immunized mice. 3 hours later, further immigration to LN was blocked with 100μg anti-CD62L. 6 hours and 24 hours after CFA, DLNs were collected, Ly5.1⁺ lymphocytes were counted by FACS. Retention rate was calculated as the ratio of cell numbers between 24 hour and 6 hour. D. WT and LIGHT^−/−mice were immunized with CFA. 2% FITC in acetone and dibutyl phthalate (v:v=1:1) was applied near the CFA site in 100μl. 6 or 24 hrs later, iLN was collagenase digested and migratory DC (FITC+CD11c+) was determined by flow cytometry. *, P < 0.05. Representative of two experiments.