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Localization of interleukin (IL) -4 but not IL-5 to human mast cell secretory granules by immunoelectron microscopy

Localization of interleukin (IL) -4 but not IL-5 to human mast cell secretory granules by immunoelectron microscopy
Localization of interleukin (IL) -4 but not IL-5 to human mast cell secretory granules by immunoelectron microscopy
BackgroundHuman mast cells synthesize and secrete many cytokines of relevance to the pathogenesis of allergic diseases such as asthma and rhinitis. In particular, interleukin (IL) ‐4 and IL‐5 are likely to play key roles in the development of the inflammatory response that characterizes these diseases. Immunohistochemical studies on human nasal and bronchial mucosal biopsies suggest that IL‐4 and IL‐5 may be stored preformed in mast cells.ObjectiveTo identify whether IL‐4 and IL‐5 are stored within mast cell secretory granules.MethodsWe used immunogold electron microscopic analysis on bronchial mucosa and lung parenchyma from resected lung specimens, and a nasal mucosal biopsy from a patient with active allergic rhinitis. Samples were fixed in 4% paraformaldehyde plus 0.5% glutaraldehyde and processed into Lowicryl K4M resin by the ‘Progressive Lowering of Temperature’ technique. Ultrathin sections were stained immunohistochemically by an indirect immunogold method.ResultsImmunoreactivity for IL‐4, but not IL‐5, was localized to the granules of mast cells in all tissue samples. IL‐5 was localized to the matrix of eosinophil granules in these samples, but neither cytokine was detected in T cells. IL‐4 immunoreactivity increased in the granules of mast cells 24 h after immunoglobulin (Ig) E‐dependent activation (mean 17.5 ± 1.4 gold particles per granule) compared with nonactivated mast cells (mean 6.8 ± 0.8 gold particles per granule, P < 0.001), suggesting replenishment of stores by newly generated protein. Immunoreactive IL‐5 remained undetectable in mast cells 24 h after activation, a time point at which they are known to secrete large quantities of this cytokine.ConclusionHuman mast cells store IL‐4 within the matrix of their granules. Very few, if any, lung or nasal mast cells store IL‐5. A store of preformed IL‐4 within mast cell granules is likely to have an important influence during the initiation and maintenance of the allergic immunological response.
electron microscopy, human mast cells, interleukin-4, interleukin-5, immunohistochemistry
0954-7894
493-500
Wilson, S.J.
21c6875d-6870-441b-ae7a-603562a646b8
Shute, J.K.
21aa47b2-7a14-4706-981d-f6cc6be58bf4
Holgate, S.T.
2e7c17a9-6796-436e-8772-1fe6d2ac5edc
Howarth, P.H.
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Bradding, P.
39990c99-3c21-44b6-b1f7-084eddf29722
Wilson, S.J.
21c6875d-6870-441b-ae7a-603562a646b8
Shute, J.K.
21aa47b2-7a14-4706-981d-f6cc6be58bf4
Holgate, S.T.
2e7c17a9-6796-436e-8772-1fe6d2ac5edc
Howarth, P.H.
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Bradding, P.
39990c99-3c21-44b6-b1f7-084eddf29722

Wilson, S.J., Shute, J.K., Holgate, S.T., Howarth, P.H. and Bradding, P. (2000) Localization of interleukin (IL) -4 but not IL-5 to human mast cell secretory granules by immunoelectron microscopy. Clinical & Experimental Allergy, 30 (4), 493-500. (doi:10.1046/j.1365-2222.2000.00756.x). (PMID:10718846)

Record type: Article

Abstract

BackgroundHuman mast cells synthesize and secrete many cytokines of relevance to the pathogenesis of allergic diseases such as asthma and rhinitis. In particular, interleukin (IL) ‐4 and IL‐5 are likely to play key roles in the development of the inflammatory response that characterizes these diseases. Immunohistochemical studies on human nasal and bronchial mucosal biopsies suggest that IL‐4 and IL‐5 may be stored preformed in mast cells.ObjectiveTo identify whether IL‐4 and IL‐5 are stored within mast cell secretory granules.MethodsWe used immunogold electron microscopic analysis on bronchial mucosa and lung parenchyma from resected lung specimens, and a nasal mucosal biopsy from a patient with active allergic rhinitis. Samples were fixed in 4% paraformaldehyde plus 0.5% glutaraldehyde and processed into Lowicryl K4M resin by the ‘Progressive Lowering of Temperature’ technique. Ultrathin sections were stained immunohistochemically by an indirect immunogold method.ResultsImmunoreactivity for IL‐4, but not IL‐5, was localized to the granules of mast cells in all tissue samples. IL‐5 was localized to the matrix of eosinophil granules in these samples, but neither cytokine was detected in T cells. IL‐4 immunoreactivity increased in the granules of mast cells 24 h after immunoglobulin (Ig) E‐dependent activation (mean 17.5 ± 1.4 gold particles per granule) compared with nonactivated mast cells (mean 6.8 ± 0.8 gold particles per granule, P < 0.001), suggesting replenishment of stores by newly generated protein. Immunoreactive IL‐5 remained undetectable in mast cells 24 h after activation, a time point at which they are known to secrete large quantities of this cytokine.ConclusionHuman mast cells store IL‐4 within the matrix of their granules. Very few, if any, lung or nasal mast cells store IL‐5. A store of preformed IL‐4 within mast cell granules is likely to have an important influence during the initiation and maintenance of the allergic immunological response.

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More information

Published date: April 2000
Keywords: electron microscopy, human mast cells, interleukin-4, interleukin-5, immunohistochemistry

Identifiers

Local EPrints ID: 190935
URI: https://eprints.soton.ac.uk/id/eprint/190935
ISSN: 0954-7894
PURE UUID: 9bfdbd62-98c3-426a-ad8c-4306378286a5

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Date deposited: 17 Jun 2011 08:30
Last modified: 23 Jul 2018 16:31

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