Figure 4.

Gp96 enhances Treg suppressive function

(A) A total of 5×10⁴ cell trace violet-labeled CD4⁺CD25^– T (Teff) cells were cultured with Tregs at a ratio of 2:1 for 3 days. Teff cell division cycle was measured using FACS (left). Suppression rate of proliferation was calculated (right).

(B–E) Rag2^−/− recipient mice were intravenously administered 4×10⁵ naïve CD4⁺ T cells (CD45.2⁺) with or without co-transfer of 4×10⁵ Treg cells (CD45.1⁺CD45.2⁺) sorted from gp96-immunized or control CD45.1⁺CD45.2⁺ mice. Experimental scheme (B). Mouse weight changes were monitored for 6 weeks (C). Hematoxylin and eosin (H&E) staining of colon at end of the experiment. Dashed lines indicate lymphocytes infiltrating areas, scale bar, 2 mm (D). Flow cytometric analysis and quantification of frequency of CD45.1 and CD45.2 cells from spleen and mesenteric lymph nodes (MLN) 6 weeks after transfer (E). Dots represent data from individual mice. The data are representative of two independent experiments with similar results. n = 5 mice/group. Mean ± SD is shown. The Student's t test or two-way ANOVA was used for statistical analysis. P < 0.05 was considered statistically significant.