![Figure 2. Eotaxin-3 is a CCR2 antagonist on monocytes. (A) Changes of [Ca++]i in fura-2–loaded cells sequentially stimulated with eotaxin-3 and MCP-1 at 90-second intervals were monitored. (B) The rate of [Ca++]i rise (% fura saturation/second) induced by MCP-1 was set to 100% and the rate after eotaxin-3 prestimulation was calculated. The results of 1 of 3 independent experiments are shown. (C) MCP-1 (•) or eotaxin-3 (▴)–induced N-acetyl-β-d-glucosaminidase release was assessed in cells treated with cytochalasin-B. Mean values (± SD) of 3 experiments performed with monocytes from different donors are shown. (D) Inhibition of MCP-1–induced N-acetyl-β-d-glucosaminidase release by eotaxin-3 was detected in monocytes sequentially stimulated with increasing concentrations of eotaxin-3 and 10 nM MCP-1. Mean values (± SD) of 3 experiments performed with monocytes from different donors are shown. (E) Monocyte chemotaxis induced by MCP-1 (10 nM) or MCP-1 (10 nM) plus eotaxin-3 (1 μM) was evaluated in Boyden microchambers. Chemotaxis index (mean ± SD) of 3 experiments performed with monocytes from different donors is shown. (F) Eotaxin-3 inhibits MCP-1–induced CCR2 internalization. The histograms represent CCR2 expression on unstimulated cells (dark-filled), cells stimulated with MCP-1 (10 nM; thin line), and cells sequentially stimulated with eotaxin-3 (1 μM) and MCP-1 (10 nM). One representative of 3 experiments performed with monocytes from different donors is shown.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/102/3/10.1182_blood-2002-09-2773/6/h81534731002.jpeg?Expires=1735840370&Signature=xbEd3PVzpJk~WQL31tFEo2Jz~mNJraZUvHya8idYVSvK~P-DqPMGmuzYJiRzHxkOKSxn1oAtVK1NsVSahpw4Qxnzy4mAcufDJgW-OTYCKzLRFaWm8~N0niqJpHmmRWUshvM90ibIn23u2RBORUrg7Q886kEDBXC5s1AAjgnZZssSgpCbumAtIRXh24~eWZHgxIBSa7Dm50~DKcjA~R1EsgiO6LDWDcv55feoE8uZ54m~7Zeb5tdAYprT~doYsxDVV4EX96bTZrlHHAfREl4XTHcadehiCVxYqOHQNOTMfSrf9g-FE~9RuCQoLz3wBK4D0cl31RdK48HekmvK3n0JSg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Eotaxin-3 is a CCR2 antagonist on monocytes. (A) Changes of [Ca++]i in fura-2–loaded cells sequentially stimulated with eotaxin-3 and MCP-1 at 90-second intervals were monitored. (B) The rate of [Ca++]i rise (% fura saturation/second) induced by MCP-1 was set to 100% and the rate after eotaxin-3 prestimulation was calculated. The results of 1 of 3 independent experiments are shown. (C) MCP-1 (•) or eotaxin-3 (▴)–induced N-acetyl-β-d-glucosaminidase release was assessed in cells treated with cytochalasin-B. Mean values (± SD) of 3 experiments performed with monocytes from different donors are shown. (D) Inhibition of MCP-1–induced N-acetyl-β-d-glucosaminidase release by eotaxin-3 was detected in monocytes sequentially stimulated with increasing concentrations of eotaxin-3 and 10 nM MCP-1. Mean values (± SD) of 3 experiments performed with monocytes from different donors are shown. (E) Monocyte chemotaxis induced by MCP-1 (10 nM) or MCP-1 (10 nM) plus eotaxin-3 (1 μM) was evaluated in Boyden microchambers. Chemotaxis index (mean ± SD) of 3 experiments performed with monocytes from different donors is shown. (F) Eotaxin-3 inhibits MCP-1–induced CCR2 internalization. The histograms represent CCR2 expression on unstimulated cells (dark-filled), cells stimulated with MCP-1 (10 nM; thin line), and cells sequentially stimulated with eotaxin-3 (1 μM) and MCP-1 (10 nM). One representative of 3 experiments performed with monocytes from different donors is shown.
