Eotaxin : a potent eosinophil chemoattractant cytokine detected in a guinea pig model of allergic airways inflammation
Eosinophil accumulation is a prominent feature of allergic inflammatory reactions, such as those occurring in the lung of the allergic asthmatic, but the endogenous chemoattractants involved have not been identified. We have investigated this in an established model of allergic inflammation, using i...
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Published in | The Journal of experimental medicine Vol. 179; no. 3; pp. 881 - 887 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
New York, NY
Rockefeller University Press
01.03.1994
The Rockefeller University Press |
Subjects | |
Online Access | Get full text |
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Abstract | Eosinophil accumulation is a prominent feature of allergic inflammatory reactions, such as those occurring in the lung of the allergic asthmatic, but the endogenous chemoattractants involved have not been identified. We have investigated this in an established model of allergic inflammation, using in vivo systems both to generate and assay relevant activity. Bronchoalveolar lavage (BAL) fluid was taken from sensitized guinea pigs at intervals after aerosol challenge with ovalbumin. BAL fluid was injected intradermally in unsensitized assay guinea pigs and the accumulation of intravenously injected 111In-eosinophils was measured. Activity was detected at 30 min after allergen challenge, peaking from 3 to 6 h and declining to low levels by 24 h. 3-h BAL fluid was purified using high performance liquid chromatography techniques in conjunction with the skin assay. Microsequencing revealed a novel protein from the C-C branch of the platelet factor 4 superfamily of chemotactic cytokines. The protein, "eotaxin," exhibits homology of 53% with human MCP-1, 44% with guinea pig MCP-1, 31% with human MIP-1 alpha, and 26% with human RANTES. Laser desorption time of flight mass analysis gave four different signals (8.15, 8.38, 8.81, and 9.03 kD), probably reflecting differential O-glycosylation. Eotaxin was highly potent, inducing substantial 111In-eosinophil accumulation at a 1-2 pmol dose in the skin, but did not induce significant 111In-neutrophil accumulation. Eotaxin was a potent stimulator of both guinea pig and human eosinophils in vitro. Human recombinant RANTES, MIP-1 alpha, and MCP-1 were all inactive in inducing 111In-eosinophil accumulation in guinea pig skin; however, evidence was obtained that eotaxin shares a binding site with RANTES on guinea pig eosinophils. This is the first description of a potent eosinophil chemoattractant cytokine generated in vivo and suggests the possibility that similar molecules may be important in the human asthmatic lung. |
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AbstractList | Eosinophil accumulation is a prominent feature of allergic inflammatory reactions, such as those occurring in the lung of the allergic asthmatic, but the endogenous chemoattractants involved have not been identified. We have investigated this in an established model of allergic inflammation, using in vivo systems both to generate and assay relevant activity. Bronchoalveolar lavage (BAL) fluid was taken from sensitized guinea pigs at intervals after aerosol challenge with ovalbumin. BAL fluid was injected intradermally in unsensitized assay guinea pigs and the accumulation of intravenously injected 111In-eosinophils was measured. Activity was detected at 30 min after allergen challenge, peaking from 3 to 6 h and declining to low levels by 24 h. 3-h BAL fluid was purified using high performance liquid chromatography techniques in conjunction with the skin assay. Microsequencing revealed a novel protein from the C-C branch of the platelet factor 4 superfamily of chemotactic cytokines. The protein, "eotaxin," exhibits homology of 53% with human MCP-1, 44% with guinea pig MCP-1, 31% with human MIP-1 alpha, and 26% with human RANTES. Laser desorption time of flight mass analysis gave four different signals (8.15, 8.38, 8.81, and 9.03 kD), probably reflecting differential O-glycosylation. Eotaxin was highly potent, inducing substantial 111In-eosinophil accumulation at a 1-2 pmol dose in the skin, but did not induce significant 111In-neutrophil accumulation. Eotaxin was a potent stimulator of both guinea pig and human eosinophils in vitro. Human recombinant RANTES, MIP-1 alpha, and MCP-1 were all inactive in inducing 111In-eosinophil accumulation in guinea pig skin; however, evidence was obtained that eotaxin shares a binding site with RANTES on guinea pig eosinophils. This is the first description of a potent eosinophil chemoattractant cytokine generated in vivo and suggests the possibility that similar molecules may be important in the human asthmatic lung. |
Author | WALSH, D. T GRIFFITHS-JOHNSON, D. A JOSE, P. J WILLIAMS, T. J COLLLINS, P. D TOTTY, N. F TRUONG, O MOQBEL, R HSUAN, J. J |
Author_xml | – sequence: 1 givenname: P. J surname: JOSE fullname: JOSE, P. J organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 2 givenname: D. A surname: GRIFFITHS-JOHNSON fullname: GRIFFITHS-JOHNSON, D. A organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 3 givenname: P. D surname: COLLLINS fullname: COLLLINS, P. D organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 4 givenname: D. T surname: WALSH fullname: WALSH, D. T organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 5 givenname: R surname: MOQBEL fullname: MOQBEL, R organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 6 givenname: N. F surname: TOTTY fullname: TOTTY, N. F organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 7 givenname: O surname: TRUONG fullname: TRUONG, O organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 8 givenname: J. J surname: HSUAN fullname: HSUAN, J. J organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom – sequence: 9 givenname: T. J surname: WILLIAMS fullname: WILLIAMS, T. J organization: National heart lung inst., dep. applied pharmacology, London SW3 6LY, United Kingdom |
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Keywords | Allergy Animal model Vertebrata Mammalia Guinea pig Rodentia Cytokine Inflammation Chemotaxis Biological activity Eosinophil Chemotactic factor |
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SubjectTerms | Amino Acid Sequence Animals Biological and medical sciences Bronchoalveolar Lavage Fluid - immunology Chemokine CCL11 Chemokine CCL4 Chemokine CCL5 Chemokines, CC Chemotaxis, Leukocyte Cytokines - biosynthesis Cytokines - chemistry Cytokines - isolation & purification Cytokines - pharmacology Disease Models, Animal Eosinophils - drug effects Eosinophils - physiology Fundamental and applied biological sciences. Psychology Fundamental immunology Guinea Pigs Humans Hypersensitivity - immunology Immediate hypersensitivity. Allergy. Anaphylaxis, etc Immunobiology Inflammation Lymphokines - chemistry Lymphokines - pharmacology Macrophage Inflammatory Proteins Male Molecular Sequence Data Monokines - chemistry Monokines - pharmacology Reaction mechanisms, antibodies, chemical mediators Recombinant Proteins - pharmacology Respiratory Tract Diseases - immunology Sequence Homology, Amino Acid |
Title | Eotaxin : a potent eosinophil chemoattractant cytokine detected in a guinea pig model of allergic airways inflammation |
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