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 (≤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 µg/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.

Keywords:
Basophils, CCR3, Chemotaxis, Histamine, Mast cells
1.
Baggiolini MB, Dewald B, Moser B: Human chemokines: An update. Annu Rev Immunol 1997;15:675–705.
2.
Mantovani A, Allavena P, Vecchi A, Sozzani S: Chemokines and chemokine receptors during activation and deactivation of monocytes and dendritic cells and in amplification of Th1 versus Th2 responses. Int J Clin Lab Res 1998;28:77–82.
3.
Luster AD: Chemokines: Chemotactic cytokines that mediate inflammation. N Engl J Med 1998;338:436–445.
4.
Kitamura M, Nakajima T, Harada S, Combadiere C, Tiffany HL, Murphy PM, Yoshie O: Molecular cloning of human eotaxin, an eosinophil-selective CC chemokine and identification of a specific eosinophil eotaxin receptor. J Biol Chem 1996;271:7725–7730.
5.
Ponath PD, Qin SX, Post TW, Wang J, Wu L, Gerard NP, Newman W, Gerard C, Mackay CR: Molecular cloning and characterization of a human eotaxin receptor expressed selectively on eosinophils. J Exp Med 1997;183:2437–2448.
6.
Daugtery BL, Siciliano SJ, DeMartino JA, Malkowitz L, Sirotina A, Springer MS: Cloning, expression and characterization of the human eosinophil eotaxin receptor. J Exp Med 1996;183:2349–2354.
7.
Uguccioni M, Mackay CR, Ochensberger B, Loetscher P, Rhis S, LaRosa GJ, Rao P, Ponath PD, Baggiolini M, Dahinden CA: High expression of the chemokine receptor CCR3 in human blood basophils. Role in activation by eotaxin, MCP-4 and other chemokines. J Clin Invest 1997;100:1137–1143.
8.
Sallusto F, Lanzavecchia A: Selective expression of the eotaxin receptor CCR3 by human T helper 2 cells. Science 1997;277:2005–2007.
9.
Gerber BO, Zanni MP, Uguccioni M, Loetscher P, Mackay CR, Pichler WJ, Yawalkar N, Baggiolini B, Moser B: Functional expression of the eotaxin receptor CCR3 in T lymphocytes co-localizing with eosinophils. Curr Biol 1997;7:836–843.
10.
Schwartz LB, Sakai K, Bradford TR, Ren S, Zweiman B, Worobec AS, Metcalfe DD: The α form of human tryptase is the predominant type present in blood at baseline in normal subjects and is elevated in those with systemic mastocytosis. J Clin Invest 1995;96:2702–2710.
11.
Uccini S, Sirianni MC, Vincenzi V, Stopacciaro A, Lesnoni LA, Parola L, Capuana M, Cerimele D, Cells M, Lanzavecchia A, Allavena P, Mantovani A, Baroni CD, Ruco LP: Kaposi’s sarcoma cells express the macrophage associated antigen mannose receptor and develop in peripheral blood cultures of Kaposi’s sarcoma patients. Am J Pathol 1997;150:929–938.
12.
de Paulis A, Marinò I, Ciccarelli A, de Crescenzo G, Concardi M, Verga L, Arbustini E, Marone G: Human synovial mast cells. I. Ultrastructural in situ and in vitro immunologic characterization. Arthritis Rheum 1996;39:1222–1233.
13.
Patella V, Casolaro V, Ciccarelli A, Pettit GR, Columbo M, Marone G: The antineoplastic bryostatins affect human basophils and mast cells differently. Blood 1995;85:1272–1281.
14.
Romagnani P, Annunziato F, Manetti R, Mavilia R, Lasagni L, Manuelli C, Vanelli GB, Maggi E, Pupilli C, Romagnani S: High CD30 ligand expression by epithelial cells and Hassal’s corpuscles in the medulla of human thymus. Blood 1998;91:3323–3332.
15.
Galli G, Annunziato F, Mavilia C, Romagnani P, Cosmi L, Manetti R, Pupilli C, Maggi E, Romagnani S: Enhanced HIV expression during Th2-oriented responses explained by the opposite regulatory effect of IL-4 and IFN-γ on fusin /CXCR4. Eur J Immunol 1998;28:3280–3290.
16.
de Paulis A, Ciccarelli A, de Crescenzo G, Cirillo R, Patella V, Marone G: Cyclosporin H is a potent and selective competitive antagonist of human basophil activation by FMLP. J Allergy Clin Immunol 1996;98:152–164.
17.
Siraganian RP: An automatic continuous-flow system for the extraction and fluorometric analysis of histamine. Anal Biochem 1974;57:383–394.
18.
Schleimer RP, Freeland HS, Peters SP, Brown KE, Perse CP: An assessment of the effects of glucocorticoids on degranulation, chemotaxis to vascular endothelial cells and formation of leucotriene B4 by purified human neutrophils. J Pharmacol Exp Ther 1992;250:598–604.
19.
Stead RH, Tomioka M, Quinonez G, Simon GT, Felten SY, Bienenstock J: Intestinal mucosal mast cells and nematode-infected rat intestines are in intimate contact with peptidergic nerves. Proc Natl Acad Sci USA 1987;84:2975–2979.
20.
Mavilia C, Scaletti C, Romagnani P, Carossino AM, Pignone A, Emmi L, Pupilli C, Pizzolo G, Maggi E, Romagnani S: Type 2 helper T (Th2) cell predominance and high CD30 expression in systemic sclerosis. Am J Pathol 1997;151:1751–1758.
21.
Irani AM, Schechter NM, Craig SS, DeBlois G, Schwartz LB: Two types of human mast cells that have distinct neutral protease compositions. Proc Natl Acad Sci USA 1986;83:4464–4468.
22.
Patella V, Marinò I, Lamparter B, Arbustini E, Adt M, Marone G: Human heart mast cells: Isolation, purification, ultrastructure, and immunologic characterization. J Immunol 1995;154:2855–2865.
© 2001 S. Karger AG, Basel
2001
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.