Macaque lymphocytes transduced by a constitutively expressed interferon beta gene display an enhanced resistance to SIVmac251 infection.
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.
Matheux
F
F
CEA, Service de Neurovirologie (DSV/DRM), CRSSA, Institut Paris Sud sur les Cytokines, Fontenay aux Roses, France.
Le Grand
R
R
Rousseau
V
V
De Maeyer
E
E
Dormont
D
D
Lauret
E
E
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D016370:Gene Products, rex / Q000032:analysis; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007111:Immunity, Cellular; D016899:Interferon-beta / Q000096:biosynthesis / Q000235:genetics*; D008214:Lymphocytes / Q000378:metabolism*; D008252:Macaca fascicularis; D051379:Mice; D008969:Molecular Sequence Data; D010641:Phenotype; D012190:Retroviridae; D017386:Sequence Homology, Amino Acid; D016097:Simian Acquired Immunodeficiency Syndrome / Q000276:immunology*; D015302:Simian Immunodeficiency Virus / Q000276:immunology; D013997:Time Factors; D014161:Transduction, Genetic
D016370:Gene Products, rex; D016899:Interferon-beta