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Recombinant retrovirus vectors for the expression of MHC class II heterodimers.
- Source :
-
Surgery today [Surg Today] 1999; Vol. 29 (6), pp. 533-41. - Publication Year :
- 1999
-
Abstract
- Class II antigens are critical in determining the fate of vascularized allografts across major histocompatibility differences. We have recently developed a new approach to induce transplantation tolerance in miniature swine by creating MHC class II antigen "molecular chimerism" in bone marrow cells of potential recipients through retrovirus-mediated gene transfer. As part of this project, the ability of a recombinant double-expression vector (ZQ32N) to express MHC class II DQA and DQB was investigated. Flow cytometry analyses of ZQ32N transfected virus-producer cells demonstrated the cell surface expression of DQa/DQb heterodimers, thus suggesting a correct transcription, translation, and transport of the swine polypeptides to the cell surface. The analyses of RNA isolated from virus particles produced from ZQ32N transfected virus-producer cells indicated the DQ sequences to be correctly packaged. However, the DQ-negative cells transduced with the ZQ32N retrovirus did not show any DQ-retrovirus surface expression. Southern and Northern blot analyses of ZQ32N transfected and transduced cells strongly suggested DNA rearrangements and deletions which could account for transgene expression loss. An analysis of transduced cell genomes suggested DNA recombinations targeted to homologous sequences within the recombinant provirus. The implications of the sequence instability in designing vectors for gene therapy of organ transplantation are discussed.
- Subjects :
- Animals
Blotting, Northern
Blotting, Southern
Flow Cytometry
Gene Transfer Techniques
HLA-DQ alpha-Chains
HLA-DQ beta-Chains
Immune Tolerance
Plasmids genetics
RNA, Viral analysis
Swine
Swine, Miniature
Transcription, Genetic
Transduction, Genetic
Transfection
DNA, Recombinant genetics
DNA, Viral genetics
Genetic Vectors genetics
HLA-DQ Antigens genetics
Retroviridae genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0941-1291
- Volume :
- 29
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Surgery today
- Publication Type :
- Academic Journal
- Accession number :
- 10385368
- Full Text :
- https://doi.org/10.1007/BF02482348