Abstracts
British Association for the Study of the Liver Meeting
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The following abstracts were presented at the British Association for the Study of the Liver Meeting, University of Southampton, UK, September 7-8, 2000.
N. Mellor1, M. Themis2, C. Selden1, M. Jones3, H. J. F. Hodgson1. 1 Centre for Hepatology, Royal Free Campus-UCL, Rowland Hill St., London, NW3 2PF.2 Cystic Fibrosis Gene Therapy Research Group, Imperial College School of Medicine, London. 3 Dept of Infectious Diseases, Imperial College School of Medicine, London. n.mellor@rfc.ucl.ac.uk
Introduction: Histidinaemia is an autosomal recessive disorder in man affecting the hepatic enzyme histidase resulting in elevated plasma and urinary histidine. A mouse model with a similar phenotype lends itself to investigating methods of gene therapy.
Aim: To revert the biochemical phenotype in by in vivo retroviral gene transfer.
Methods: A cDNA encoding normal histidase was cloned into the LNCX retroviral construct under the control of the CMV promoter. Virus was concentrated and administered to histidinaemic mice primed for hepatocyte transfection by partial hepatectomy. HPLC flow scintillation was used to assess in vivo catalysis of 14C-histidine. Hepatic cytosol was examined for histidase expression by Western analysis and enzyme activity by conversion of histidine to urocanic acid.
Results: Infected animals showed a
33%±15% enhancement of metabolism as shown
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