Macaque Lymphocytes Transduced by a Constitutively Expressed Interferon beta Gene Display an Enhanced Resistance to SIVmac251 Infection

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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 PCRamplified and sequenced the Ma IFN-beta-coding sequence, and placed it under the control of a PstI-Nru I 0.6-kb fragment of the murine H-2Kb gene promoter in the MFG-KbMaIFN beta 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-KbMaIFN beta 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.