This Article Full Text Full Text (PDF) All Versions of this Article: 2005-0377OCv1 35/1/72    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Davidson, H. Articles by Penque, D. Search for Related Content PubMed PubMed Citation Articles by Davidson, H. Articles by Penque, D.

Human-Specific Cystic Fibrosis Transmembrane Conductance Regulator Antibodies Detect In Vivo Gene Transfer to Ovine Airways

Medical Sciences (Medical Genetics), University of Edinburgh, Western General Hospital, Edinburgh; The Wellcome Trust Centre in Comparative Respiratory Medicine, Easter Bush Veterinary Centre, University of Edinburgh, Roslin; Gene Medicine Group, Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford; UK Cystic Fibrosis Gene Therapy Consortium, London, Oxford, and Edinburgh, United Kingdom; Centro de Genética Humana, Instituto Nacional Saúde Dr Ricardo Jorge; and Department of Chemistry and Biochemistry, Faculty of Sciences, University of Lisbon, Lisbon, Portugal

Correspondence and requests for reprints should be addressed to Heather Davidson, Medical Sciences (Medical Genetics), University of Edinburgh, Molecular Medicine Centre, Western General Hospital, Edinburgh EH4 2XU, United Kingdom. E-mail: H.Davidson{at}ed.ac.uk

A panel of 11 human cystic fibrosis transmembrane conductance regulator (hCFTR) antibodies were tested in ovine nasal, tracheal, and bronchial epithelial brushings. Two of these, G449 (polyclonal) and MATG1104 (monoclonal), recognized hCFTR but did not cross react with endogenous sheep CFTR. This specificity allows immunologic detection of hCFTR expressed in gene transfer studies in sheep against the background of endogenous ovine CFTR, thus enhancing the value of the sheep as a model animal in which to study CFTR gene transfer. Studies on mixed populations of human and sheep nasal epithelial cells showed that detection of hCFTR by these two antibodies was possible even at the lowest proportion of human cells (1:100). The hCFTR gene was delivered in vivo by local instillation using polyethylenimine-mediated gene transfer to the ventral surface of the ovine trachea and hCFTR mRNA and protein levels scored in a blinded fashion. Despite abundant hCFTR mRNA expression, the number of cells expressing hCFTR protein detectable by G449 was low ( 0.006–0.05%). Immunohistochemistry for hCFTR in animals treated by whole-lung aerosol demonstrated positive cells in sections of tracheal epithelium and in distal conducting airways. The strategic use of hCFTR-specific antibodies supports the utility of the normal sheep as a model for hCFTR gene transfer studies.

Key Words: CFTR antibodies • cystic fibrosis • gene transfer • nasal brushing cells • sheep model