Figure 6.

Hepatocytes Captured Treg Cells More Efficiently than Non-Treg Cells and Selectively Acidified Treg Cell-Containing Vesicles

(A) CD4⁺ T cells were isolated from peripheral blood (BMQC, blue) and co-cultured overnight with a confluent monolayer of Huh-7 cells (CMFDA, green). The cells were fixed and stained for Foxp3 to detect T regulatory (Treg) cells (red, anti-mouse immunoglobulin G [IgG]-Alexa 594). Approximately 60% of captured T cells were Foxp3⁺. Foxp3⁻ T cells were labeled more brightly with BMQC and easy to distinguish.

(B) Treg and non-Treg cells were isolated from peripheral blood using magnetic beads and added to wells containing Huh-7 cells at equal numbers. 3 h later, non-associating T cells were washed off, and internalized cells were enumerated by confocal microscopy. Each dot represents average internalized cells per 100 Huh-7 cells in independent experiments.

(C) Treg and non-Treg cells were isolated from peripheral blood and co-cultured overnight with Huh-7 cells in the presence of colorless pHrodo Red dextran, which fluoresces under low-pH conditions. Bright acidic vesicles were enumerated in random fields of view and plotted for three independent experiments.

Error bars show SD. ^∗∗∗∗p < 0.0001. Statistics were derived from an unpaired, two-tailed Student’s t test.