Macaque Lymphocytes Transduced by a Constitutively Expressed Interferon beta Gene Display an Enhanced Resistance to SIVmac251 Infection
| Autor: | Edward De Maeyer, Véronique Rousseau, Dominique Dormont, Roger Le Grand, Evelyne Lauret, F Matheux |
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| Rok vydání: | 1999 |
| Předmět: |
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 |
| Zdroj: | Human Gene Therapy. 10:429-440 |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/10430349950018878 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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