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Expression of a swine class II gene in murine bone marrow hematopoietic cells by retroviral-mediated gene transfer.

Shafer, GE; Emery, DW; Gustafsson, K; Germana, S; Anderson, WF; Sachs, DH; LeGuern, C; (1991) Expression of a swine class II gene in murine bone marrow hematopoietic cells by retroviral-mediated gene transfer. Proc Natl Acad Sci U S A , 88 (21) pp. 9760-9764.

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Abstract

As a first step in assessing the efficacy of a gene transfer approach to the induction of transplantation tolerance in our miniature swine model, double-copy retroviral vectors engineered to express a drug-resistance marker (neomycin) and a swine class II DRB cDNA were constructed. Infectious particles containing these vectors were produced at a titer of greater than 1 x 10(6) G418-resistant colony-forming units/ml using both ecotropic and amphotropic packaging cell lines. Flow cytometric analysis of DRA-transfected murine fibroblasts subsequently transduced with virus-containing supernatants demonstrated that the transferred sequences were sufficient to produce DR surface expression. Cocultivation of murine bone marrow with high-titer producer lines leads to the transduction of 40% of granulocyte/macrophage colony-forming units (CFU-GM) as determined by the frequency of colony formation under G418 selection. After nearly 5 weeks in long-term bone marrow culture, virus-exposed marrow still contained G418-resistant CFU-GM at a frequency of 25%. In addition, virtually all of the transduced and selected colonies contained DRB-specific transcripts. These results suggest that a significant proportion of very primitive myelopoietic precursor cells can be transduced with the DRB recombinant vector and that vector sequences are expressed in the differentiated progeny of these cells.

Type: Article
Title: Expression of a swine class II gene in murine bone marrow hematopoietic cells by retroviral-mediated gene transfer.
Location: United States
Keywords: Animals, Bone Marrow Cells, Cloning, Molecular, Gene Expression, Genes, MHC Class II, Genetic Vectors, Hematopoietic Stem Cells, Mice, RNA, Messenger, Retroviridae, Swine, Transcription, Genetic, Transfection
UCL classification: UCL > School of Life and Medical Sciences
UCL > School of Life and Medical Sciences > Faculty of Population Health Sciences > Institute of Child Health
URI: http://discovery.ucl.ac.uk/id/eprint/1354539
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