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Macaque Lymphocytes Transduced by a Constitutively Expressed Interferon beta Gene Display an Enhanced Resistance to SIVmac251 Infection
- Source :
- Human Gene Therapy. 10:429-440
- Publication Year :
- 1999
- Publisher :
- Mary Ann Liebert Inc, 1999.
-
Abstract
- We are developing a method of gene therapy of HIV infection based on the low constitutive expression of an interferon beta (IFN-beta) gene in HIV target cells. Herein we report the first step in the development of a relevant animal model, provided by the macaque (Macaca fascicularis) infected with a pathogenic SIVmac251 isolate. To avoid the possibility of in vivo rejection of macaque lymphocytes expressing Hu IFN-beta, we have PCR-amplified and sequenced the Ma IFN-beta-coding sequence, and placed it under the control of a PstI-NruI 0.6-kb fragment of the murine H-2Kb gene promoter in the MFG-K(b)MaIFNbeta retroviral vector. Lymphocytic CEMX174 cells, transduced by coculture on packaging cells with this construct, harbored a mean of 0.07 to 1.2 copies of the IFN-beta transgene per cell, and were characterized by an IFN production ranging from 75 to 750 units per 5 x 10(5) cells per 3 days. The IFN-beta-transduced populations displayed an enhanced resistance against the pathogenic SIVmac251 isolate. Control experiments showed that the enhanced resistance could not be ascribed to the Ma IFN-beta released during the 3 days of coculture by the packaging cells, or to the mere transduction with a retroviral vector. Macaque lymphocytes transduced by the MFG-K(b)MaIFNbeta retroviral vector by coculture on packaging cells, acquired a mean number of IFN-beta transgene copies per cell ranging from 0.03 to 0.1. Such transduction led to the release of IFN-beta into the culture medium, ranging from 10 to 20 units per 5 x 10(5) cells per 3 days. This increased the anti-SIV resistance of the lymphocytes, as demonstrated by a decreased p27 antigen release into the culture medium, without affecting lymphocyte proliferation.
- Subjects :
- Time Factors
Transgene
Genetic enhancement
Genetic Vectors
Molecular Sequence Data
Simian Acquired Immunodeficiency Syndrome
Lymphocyte proliferation
Macaque
3T3 cells
Viral vector
Mice
Transduction (genetics)
Transduction, Genetic
biology.animal
Genetics
medicine
Animals
Humans
Lymphocytes
Molecular Biology
Gene
Immunity, Cellular
Base Sequence
Dose-Response Relationship, Drug
Sequence Homology, Amino Acid
biology
3T3 Cells
Genetic Therapy
Interferon-beta
Virology
Gene Products, rex
Macaca fascicularis
Phenotype
Retroviridae
medicine.anatomical_structure
Molecular Medicine
Simian Immunodeficiency Virus
Cell Division
Subjects
Details
- ISSN :
- 15577422 and 10430342
- Volume :
- 10
- Database :
- OpenAIRE
- Journal :
- Human Gene Therapy
- Accession number :
- edsair.doi.dedup.....6c8d588a623367ebfe17fedccb68cee9