Human-specific cystic fibrosis transmembrane conductance regulator antibodies detect in vivo gene transfer to ovine airways

H. Davidson, G. McLachlan, A. Wilson, A. C. Boyd, A. Doherty, G. MacGregor, L. Davies, H. A. Painter, R. Coles, S. C. Hyde, D. R. Gill, M. D. Amaral, D. D. Collie, D. J. Porteous, D. Penque

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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 (approximately 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.
Original languageEnglish
Pages (from-to)72-83
Number of pages12
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Issue number1
Publication statusPublished - 2006

Keywords / Materials (for Non-textual outputs)

  • Amino Acid Sequence Animals Antibodies/*immunology Antibody Specificity Cell Count Cystic Fibrosis Transmembrane Conductance Regulator/chemistry/*immunology Epithelial Cells/cytology Female Gene Expression *Gene Transfer Techniques Humans Mice Mice, Inbred BALB C Molecular Sequence Data RNA, Messenger/genetics/metabolism Respiratory System/*metabolism Sequence Alignment Sheep/*metabolism Species Specificity


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