β-chemokine expression and distribution in paraffin-embedded transplant renal biopsy sections: analysis by scanning laser confocal microscopy |
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Authors: | H Robertson Janice Wheeler Adrian R Morley Trevor A Booth David Talbot John A Kirby |
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Institution: | (1) Department of Pathology, Royal Victoria Infirmary, Queen Victoria Road, Newcastle upon Tyne NE1 4LP, UK e-mail helen.robertson@ncl.ac.uk Tel. +44 191 222 6000 ext. 7174; fax +44 191 222 8100, GB;(2) Public Health Laboratory, Newcastle General Hospital, Newcastle upon Tyne, UK, GB;(3) Department of Pathology, Royal Victoria Infirmary, Newcastle upon Tyne NE1 4LP, UK, GB;(4) Biomedical Electron Microscopy, Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK, GB;(5) Department of Surgery, Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, UK, GB |
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Abstract: | Previous immunohistochemical and in situ hybridisation studies have shown that, in tubulitis associated with acute cellular
rejection of human renal allografts, intratubular T cells proliferate and are fully activated in situ. In the immunohistochemical
study reported here we have attempted to establish some understanding of the involvement of the β-chemokines RANTES, MCP-1,
MIP-1α and MIP-1β in recruiting T cells to the intratubular site. Paraffin-embedded routine biopsy sections were treated for
conventional indirect immunofluorescence to detect the selected chemokines. Scanning laser confocal microscopy was used to
provide a measure of fluorescence intensity resulting from binding of FITC-labelled secondary antibody. Cells expressing chemokines
could be identified and, within the limits of the staining method, it was possible to obtain a semi-quantitative assessment
of individual chemokine activity at different points in biopsy sections by constructing a profile of fluorescence intensity.
High concentrations of chemokines (especially RANTES, MIP-1β and/or MIP-1α) were localised to the basolateral surface of tubular
epithelial cells (TEC). MCP-1 was also consistently present but at a lower level than RANTES except in one case identified
as BANFF category 3. There was diffuse distribution of chemokines in the interstitial matrix and low intensity fluorescence
outlined some endothelial cells of peritubular venules and interstitial fibroblast-like cells. Our results suggest a mechanism
for specific chemotactic recruitment of inflammatory cells by TEC-produced chemokines.
Accepted: 22 January 1998 |
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