title: Combined Antiviral Therapy Using Designed Molecular Scaffolds Targeting Two Distinct Viral Functions, HIV-1 Genome Integration and Capsid Assembly.
abstract: Designed molecular scaffolds have been proposed as alternative therapeutic agents against HIV-1. The ankyrin repeat protein (Ank(GAG)1D4) and the zinc finger protein (2LTRZFP) have recently been characterized as intracellular antivirals, but these molecules, used individually, do not completely block HIV-1 replication and propagation. The capsid-binder Ank(GAG)1D4, which inhibits HIV-1 assembly, does not prevent the genome integration of newly incoming viruses. 2LTRZFP, designed to target the 2-LTR-circle junction of HIV-1 cDNA and block HIV-1 integration, would have no antiviral effect on HIV-1-infected cells. However, simultaneous expression of these two molecules should combine the advantage of preventive and curative treatments. To test this hypothesis, the genes encoding the N-myristoylated Myr(+)Ank(GAG)1D4 protein and the 2LTRZFP were introduced into human T-cells, using a third-generation lentiviral vector. SupT1 cells stably expressing 2LTRZFP alone or with Myr(+)Ank(GAG)1D4 showed a complete resistance to HIV-1 in viral challenge. Administration of the Myr(+)Ank(GAG)1D4 vector to HIV-1-preinfected SupT1 cells resulted in a significant antiviral effect. Resistance to viral infection was also observed in primary human CD4+ T-cells stably expressing Myr(+)Ank(GAG)1D4, and challenged with HIV-1, SIVmac, or SHIV. Our data suggest that our two anti-HIV-1 molecular scaffold prototypes are promising antiviral agents for anti-HIV-1 gene therapy.
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.|Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Wannisa
authors: W
authors: Khamaikawin
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.|Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Somphot
authors: S
authors: Saoin
authors: Division of Clinical Immunology and Transfusion Sciences, School of Allied Health Sciences, University of Phayao, Phayao, Thailand.
authors: Sawitree
authors: S
authors: Nangola
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.|Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Koollawat
authors: K
authors: Chupradit
authors: Research Institute for Health Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Supachai
authors: S
authors: Sakkhachornphop
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.|Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Sudarat
authors: S
authors: Hadpech
authors: Division of Instruments for Research, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors: Nattawat
authors: N
authors: Onlamoon
authors: Department of Pathology & Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
authors: Aftab A
authors: AA
authors: Ansari
authors: Department of Pathology & Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
authors: Siddappa N
authors: SN
authors: Byrareddy
authors: University Lyon 1 & INRA UMR-754, Retrovirus and Comparative Pathology, Lyon, France.
authors: Pierre
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authors: Boulanger
authors: University Lyon 1 & INRA UMR-754, Retrovirus and Comparative Pathology, Lyon, France.
authors: Saw-See
authors: SS
authors: Hong
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA.
authors: Bruce E
authors: BE
authors: Torbett
authors: Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.|Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Chatchai
authors: C
authors: Tayapiwatana