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. Author manuscript; available in PMC: 2008 Mar 17.

Published in final edited form as: Cell Immunol. 2007 Jan 25;243(1):19–29. doi: 10.1016/j.cellimm.2006.12.001

CCL5-induced down-regulation of CCR5 on PBMCs. 10⁷ cells/mL were incubated without chemokine or with 100 ng/mL CCL5 for 1 hour at 4°C or 37°C. Approximately 10⁶ cells were stained with anti-CD14, anti-CD3 and anti-CCR5 antibodies to determine the effects of chronic CCL5 exposure on receptor expression on non-migrating monocytes and T-cells. Both T-cells (Figure 1a) and monocytes (Figure 1b) showed reduced CCR5 expression when incubated with CCL5 at 37°C relative to input cells and cells incubated without CCL5. There was less reduction seen on both subpopulations of mononuclear cells incubated with CCL5 at 4°C, indicating that reduced CCR5 expression was secondary to receptor internalization. Shown are representative overlays from four separate experiments using two different donors. The vertical hashed line represents the gating index for these experiments based on the isotype controls.

CCL5-induced down-regulation of CCR5 on PBMCs. 10⁷ cells/mL were incubated without chemokine or with 100 ng/mL CCL5 for 1 hour at 4°C or 37°C. Approximately 10⁶ cells were stained with anti-CD14, anti-CD3 and anti-CCR5 antibodies to determine the effects of chronic CCL5 exposure on receptor expression on non-migrating monocytes and T-cells. Both T-cells (Figure 1a) and monocytes (Figure 1b) showed reduced CCR5 expression when incubated with CCL5 at 37°C relative to input cells and cells incubated without CCL5. There was less reduction seen on both subpopulations of mononuclear cells incubated with CCL5 at 4°C, indicating that reduced CCR5 expression was secondary to receptor internalization. Shown are representative overlays from four separate experiments using two different donors. The vertical hashed line represents the gating index for these experiments based on the isotype controls.