Anti rat cd25

All the fluorescently labelled cells were analysed using a FACScalibur. The following antibodies were used in flow cytometric analysis of the activation of inflammatory cells in blood and the spleen from the recipient rats: Rat CD45 HRZN V450, Rat Granulocytes RP-1, Rat CD8a PE MAB, Rat CD11b APC MAB (BD PMG, Franklin, NJ, USA), and Anti-Rat MHC Class II (eBioscience, San Diego, CA, USA) were used to monitor rat monocytes (Figure 2A). Anti-CD3 and anti-CD16 (BD PMG) were used to detect rat natural killer (NK) cells (Figure 2B). Anti-rat CD3, anti-CD4 (BD PMG), anti-CD25, and anti-Foxp3 (eBioscience, USA) were used to monitor rat CD4⁺CD25⁺Foxp3⁺ regulatory T cells (Tregs) (Figure 2C). Rat CD4⁺ T cells were analysed using Rat CD45 HRZN V450, Rat CD3 FITC MAB, Rat CD4 PE-CY7 MAB, (BD PMG, USA)and Anti-Rat CD25 (eBioscience, USA); CD8⁺ T cells were measured using Rat CD45 HRZN V450, Rat CD3 FITC MAB, Rat CD8a PE MAB, (BD PMG, USA) and Anti-Rat CD25 (eBioscience, USA) (Figure 2D).Figure 2

Analysis process of inflammatory cells. (A) CD45 ​+ ​RP1 ​+ ​CD8 ​+ ​CD11b ​+ ​MHC II → monocytes; (B) CD3 ​+ ​CD16 → NK cells; (C) CD25 ​+ ​Foxp3 ​+ ​CD3 ​+ ​CD4 → Tregs; (D) CD45 ​+ ​CD3 ​+ ​CD4 ​+ ​CD25 → CD4⁺ T cells and CD8⁺ T cells. NK = natural killer; Tregs = regulatory T cells.

Figure 2

Absolute and relative numbers of lymphocytes of various subpopulations in peripheral blood were counted using flow cytometry (Beckman Coulter). The following antibodies (eBioscience) were used for immune phenotypic analysis of the main subpopulations of lymphocytes: anti-rat CD3 (for T lymphocytes); anti-rat CD4 (for T helpers); anti-rat CD8a (for cytotoxic T cells); anti-rat CD45R (for B lymphocytes); anti-rat CD25 (for activated T cells); anti-mouse/rat Foxp3 (for regulatory T cells); and anti-rat CD314 (for NK cells). Erythrocytes were lysed with OptiLyse C solution (eBioscience).