Regulatory T cells can migrate to follicles upon T cell activation and suppress GC-Th cells and GC-Th cell–driven B cell responses
J. Clin. Invest. Hyung W. Lim, et al. 114:1640 doi:10.1172/JCI22325 [Go to this article.]

Figure 4
Chemotaxis assay. (A) Chemotactic behaviors of CD4⁺CD25⁺CD69^– Tregs and CD4⁺CD25⁺CD69⁺ T cells. (B) CD4⁺CD25⁺CD69^– Tregs poorly migrate to CCL1. Fresh tonsil mononuclear cells were used as input cells for chemotaxis assays. Indicated chemokines were first titrated to determine optimal concentrations: CXCL13 (4,000 ng/ml), CXCL12 (100 ng/ml), CXCL10 (1,000 ng/ml), CCL19 (2,000 ng/ml), CCL17 (200 ng/ml), CCL4 (100 ng/ml), and CCL1 (500 ng/ml). Cells were allowed to migrate for 3 hours. The migrated cells and input cells were harvested, stained for CD4, CD25, and CD69, and counted by a FACSCalibur. Specific migration after subtraction of the background migration is shown. The background migration rates (percent averages and SEM, 3 experiments) for the 4 subsets were 25 ± 2.6 (CD4⁺CD25^–CD69⁺), 13.4 ± 4.3 (CD4⁺CD25⁺CD69⁺), 6.1 ± 0.4 (CD4⁺CD25⁺CD69^– Treg), and 7.2 ± 2 (CD4⁺CD25^–CD69^–). The averages and SEM of the data obtained from 3 (A) and 4 (B) independent experiments are shown. *Significant differences between the 2 subsets (A) or from CD4⁺CD25⁺CD69^– Tregs (B). The P values were 0.048 (CXCL13) and 0.015 (CCL19) in A; and 0.046 (CD25⁺CD69^– Treg vs. CD25⁺CD69⁺), 0.007 (CD25⁺CD69^– Treg vs. CD25^–CD69⁺), and 0.03 (CD25⁺CD69^– Treg vs. CD25^–CD69^–) in B.