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Characterization of the ovine ortholog of secretory leukoprotease inhibitor

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Abstract

There is great interest in the use of the sheep as a model for the investigation of inflammation in the lung. The serine antiproteases secretory leukoprotease inhibitor (SLPI) and elafin are important “alarm antiproteases” in the lung and have potentially important roles in the innate immune response. SLPI was first characterized in man and subsequently in murine, porcine, and rat tissues. Here we present the first data concerning the gene and cDNA sequence encoding for the ovine ortholog of SLPI, a protein of 132 amino acids with 66% sequence identity at the amino acid level with human SLPI. A 24-amino-acid signal sequence signifies that, like the other mammalian ortholoags, ovine SLPI is a secreted protein. Tissue distribution of expression is demonstrated by reverse transcription polymerase chain reaction (RT-PCR) and shows features similar to SLPI expression in other mammals, specifically at mucosal surfaces such as the upper respiratory and intestinal tracts, and also the skin, liver, and kidney. This distribution lends credence to SLPI having important roles in innate immunity. We have also cloned the ovine SLPI cDNA into an expression vector and expressed the ovine SLPI protein in vitro. This has enabled us to demonstrate that ovine SLPI is correctly processed (Western blot analysis and SELDI-TOF mass spectrometry analysis) and has biological antihuman neutrophil elastase activity. In summary, the ovine ortholog of SLPI shows similarities to other members of the SLPI family and has all the features of a modulator of innate immunity.

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Acknowledgments

We are grateful to the Wellcome Trust for funding this work (Clinical Research Training Award in Veterinary Medicine for T. Brown) and to M. Marsden for expert technical assistance. The MRC and the Cystic Fibrosis Trust are also acknowledged for funding the SELDI-related experimental work. We also thank Prof. K. Johnson (Molecular Genetics, Anderson College, Institute of Biomedical Sciences, University of Glasgow, UK) and Dr. T. D. Tetley (Lung Cell Biology, National Heart and Lung Institute, Imperial College, London, UK) for their help and input in this project.

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Authors and Affiliations

  1. Rayne Laboratory, Respiratory Medicine Unit, MRC Centre for Inflammation Research, University of Edinburgh, Edinburgh, Scotland, EHS 9AG, UK

    Thomas I. Brown & Jean-Michel Sallenave

  2. Wellcome Trust Centre for Research in Comparative Respiratory Medicine, The University of Edinburgh, Easter Bush Veterinary Centre, Roslin Midlothian, Scotland, UK

    Thomas I. Brown & David D. Collie

  3. Molecular Genetics, Anderson College, Institute of Biomedical Sciences, University of Glasgow, Glasgow, Scotland, UK

    Rohit Mistry

  4. Molecular Medicine Centre, Western General Hospital, Edinburgh, Scotland, UK

    Robert Gray & Margaret Imrie

Authors
  1. Thomas I. Brown
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  2. Rohit Mistry
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  3. Robert Gray
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  4. Margaret Imrie
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  5. David D. Collie
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  6. Jean-Michel Sallenave
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Correspondence to Jean-Michel Sallenave.

Additional information

Abbreviations:SLPI: secretory leukoprotease inhibitor; BAC: bacterial artificial chromosome; RACE: rapid amplification of cDNA ends; WAP: whey acidic protein; REST: rapidly evolving seminal vesicle transcribed; TOM: trappin ovine molecule; HNE: human neutrophil elastase; EIC: elastase inhibitory capacity; PAGE: polyacrylamide gel electrophoresis; UTR: untranslated region ; SELDI-TOF: surface-enhanced laser desorption/ionization time of flight.

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Brown, T.I., Mistry, R., Gray, R. et al. Characterization of the ovine ortholog of secretory leukoprotease inhibitor. Mamm Genome 16, 621–630 (2005). https://doi.org/10.1007/s00335-005-0030-2

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  • DOI: https://doi.org/10.1007/s00335-005-0030-2

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