Production of glycosylated physiologically 'normal' human α1-antitrypsin by mouse fibroblasts modified by insertion of a human α1-antitrypsin cDNA using a retroviral vector

R. I. Garver, A. Chytil, S. Karlsson, G. A. Fells, M. L. Brantly, M. Courtney, P. W. Kantoff, A. W. Nienhuis, W. F. Anderson, R. G. Crystal

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α1-Antitrypsin (α1AT) deficiency is a hereditary disorder characterized by reduced serum levels of α1AT, resulting in destruction of the lower respiratory tract by neutrophil elastase. As an approach to augment α1AT levels in this disorder with physiologically normal human α1AT, we have integrated a full-length normal human α1AT cDNA into the genome of mouse fibroblasts. To accomplish this, the retroviral vector N2 was modified by inserting the simian virus 40 early promoter followed by the α1AT cDNA. Southern analysis demonstrated that the intact cDNA was present in the genome of selected clones of the transfected murine fibroblasts ψ2 and infected NIH 3T3. The clones produced three mRNA transcripts (5.8, 4.8, and 2.4 kilobases) containing human α1AT sequences, secreted an α1AT molecule recognized by an anti-human α1AT antibody, with the same molecular mass (52 kDa) as normal human α1AT and that complexed with and inhibited human neutrophil elastase. The ψ2 produced α1AT was glycosylated, and when infused intravenously into mice, it had a serum half-life similar to normal α1AT purified from human plasma and markedly longer than that of nonglycosylated human α1AT cDNA-directed yeast-produced α1AT. These studies demonstrate the feasibility of using a retroviral vector to insert the normal human α1AT cDNA into non-α1AT-producing cells, resulting in the synthesis and secretion of physiologically 'normal' human α1AT.

Original languageEnglish
Pages (from-to)1050-1054
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number4
Publication statusPublished - 8 May 1987


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