Novel Process of Intrathymic Tumor-Immune Tolerance through CCR2-Mediated Recruitment of Sirpα+ Dendritic Cells: A Murine Model (original) (raw)
Figure 3
Antigen-specific Treg generation by Sirpα+ cDCs in collaboration with IL-2 in a physiological condition.
(A) At 2 days after the last intravenous injection of OVA protein (2 mg) into DO11.10 mice, Foxp3 and CD25 expression on DO11.10high (R1) thymocytes was examined. PBS was injected as a control. Percentages of CD25highFoxp3−, CD25highFoxp3+, and CD25lowFoxp3+ cells are shown in each panel. Representative results from three independent experiments are shown**.** (B) Expression of CD25 and Foxp3 on DO11.10high mature thymocytes after stimulation with IL-2 in vitro. Percentages of CD25highFoxp3+ and CD25lowFoxp3+ cells are shown in each panel. Representative results from three independent experiments are shown. (C) At 2 days after twice intravenous injection of OVA protein, thymocytes were cultured with 2 ng/ml IL-2 for 24 hrs in the presence or absence of each blocking Ab. Percentage of CD25highFoxp3+ cells was determined. Data represent mean ± SD from three independent experiments. *, p<0.01. (D) Percentage of CD25highFoxp3+ cells among DO11.10high thymocytes in BMC mice was analyzed at 2 days after intravenous injection with OVA protein. Gray-filled symbol represents OVA protein-injected mice. Data from non-BMC DO11.10 mouse are shown as a symbol of 100% chimerism of DO11.10 thymocytes. PBS (unfilled circle) and BSA (unfilled triangle) were injected as controls. Percentage of chimerism = % of DO11.10high thymocytes in BMC thymus/% of DO11.10high thymocytes in non-BMC DO11.10 thymus x 100. Each symbol indicates one animal.