Stereoscopic antiviral protection by 3D8 scFv antibody as a DNase in nucleus and a RNase in cytoplasm to DNA virus infection on HeLa cells : Temporal and spatial protection

Abstract

3D8 scFv is a recombinant protein of anti-DNA monoclonal antibody isolated from an autoimmune-prone MRL mice. In the previous experiments, 3D8 scFv was able to exhibit degradation of both DNA and RNA with the hydrolyzing catalytic activity. In this study, we demonstrated that 3D8 scFv transfected HeLa cell lines also have the antiviral effects against virus infection. Stable 3D8 scFv cell lines were identified by qRT-PCR, FACS and confocal microscopy. The 3D8 scFv cell lines were then challenged with two DNA viruses, Herpes simplex virus-1 (HSV-1) and pseudorabies virus (PRV). HeLa cell lines expressing 3D8 scFv conferred the resistance to HSV-1 and PRV by plaque forming assay, CPE, and immunoblotting. Also, FRET assay and qRT-PCR analysis were accomplished for identifying nuclease activity on normal cell and 3D8 scFv stable cell lines to verify the hydrolysis of virus gene during the immediately early stage of virus infection process. This results demonstrated that the host genome is unaffected by 3D8 scFv and the DNA virus is thought to be degraded by the DNase activity of the nuclear 3D8 scFv primarily, and any viral transcript escaped from this degradation could be subjected to the digestion by the RNase activity of the cytosolic 3D8 scFv, which was supported by our experimental data. In conclusion, 3D8 scFv expressing HeLa cells have a novel antiviral mechanism by nuclease hydroly