Expression of the chemokine receptor CCR3 on human mast cells.
BACKGROUND
The aim of this study was to investigate whether human mast cells express functional active CCR3 receptors, which are activated by CC chemokines. These ligands include the CCR3-selective chemokines eotaxin and eotaxin-2 and the more promiscuous CC chemokines, MCP-4, MCP-3, MCP-2 and RANTES.


METHODS
Immunohistochemical analysis was performed on skin, gut and lung specimens. Double immunostaining was performed with anti-CCR3 and antitryptase, and anti-CCR3 and antichymase antibody (Ab) by using the avidin-biotin-peroxidase system with two different substrates. Mast cells were isolated and purified from human lung parenchyma (HLMC) by countercurrent elutriation followed by discontinuous Percoll density gradient. Flow-cytometric analysis of HLMC surface CCR3 expression was performed with the monoclonal Ab anti-CCR3 (7B11). Functional activation of HLMC was verified by the ability of cells to release histamine and/or migrate in response to eotaxin.


RESULTS
High percentages (>70%) of tryptase-positive cells showing CCR3 expression were found in the skin and in the intestinal submucosa, whereas much lower percentages (< or = 20%) were found in the intestinal mucosa and in the lung interstitium. Eotaxin (1-100 nM) neither induced histamine release from HLMC nor enhanced anti-IgE-induced histamine release. In contrast, eotaxin (10-100 nM) and RANTES (10-100 nM) induced HLMC chemotaxis in vitro. Preincubation of HLMC with antibody anti-CCR3 (5 microg/ml) before loading into the chemotaxis chamber abrogated chemotaxis elicited by eotaxin. Double immunostaining with anti-CCR3 and anti-chymase antibody showed that the vast majority of CCR3-expressing mast cells in the various human tissues examined were tryptase-chymase double-positive.


CONCLUSIONS
These results indicate that CCR3 is expressed on human mast cells and that these cells are attracted by CCR3-binding chemokines.
de Paulis
A
A
Division of Clinical Immunology and Allergy, University of Naples Federico II, Naples, Italy.
Annunziato
F
F
Di Gioia
L
L
Romagnani
S
S
Carfora
M
M
Beltrame
C
C
Marone
G
G
Romagnani
P
P
D002478:Cells, Cultured; D054413:Chemokine CCL11; D018946:Chemokine CCL5 / Q000494:pharmacology; D019742:Chemokines, CC; D002633:Chemotaxis; D016207:Cytokines / Q000494:pharmacology; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000378:metabolism; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D054397:Receptors, CCR3; D019707:Receptors, Chemokine / Q000378:metabolism*; D012867:Skin / Q000378:metabolism
C515456:CCL11 protein, human; C515350:CCR3 protein, human; D054413:Chemokine CCL11; D018946:Chemokine CCL5; D019742:Chemokines, CC; D016207:Cytokines; D054397:Receptors, CCR3; D019707:Receptors, Chemokine