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(American Journal of Pathology. 2004;165:1163-1173.)
© 2004 American Society for Investigative Pathology

Inhibition of Hepatitis C Virus Nonstructural Protein, Helicase Activity, and Viral Replication by a Recombinant Human Antibody Clone

Ramesh Prabhu*, Nutan Khalap*, Roberto Burioni{dagger}, Massimo Clementi{ddagger}, Robert F. Garry§ and Srikanta Dash*

From the Departments of Pathology and Laboratory Medicine* and Microbiology,§ Tulane University Health Sciences Center, New Orleans, Louisiana; the Istituto di Microbiologia Facoltà di Medicina e Chirurgia,{dagger} Università di Ancona, Ancona, Italy; and the Università "Vita-Salute San Raffaele,"{ddagger} Instituto di Ricovero e Cura a Carattera Scientifico San Raffaele, Milan, Italy

Hepatitis C virus (HCV) nonstructural protein 3 (NS3), with its protease, helicase, and NTPase enzymatic activities, plays a crucial role in viral replication, and therefore represents an ideal target for the development of anti-viral agents. We have developed a recombinant human antibody (Fab) that reacts with the helicase domain of HCV NS3. The affinity-purified Fab antibody completely inhibited the helicase activity of HCV NS3 at equimolar concentration. To evaluate the effect of the Fab on HCV replication, the clone encoding the Fab gene was put into an expression vector, which converts Fab into a complete IgG1 antibody. Using a DNA-based transfection model, we demonstrated that intracellular expression of this antibody resulted in significant reduction of HCV-negative strand RNA synthesis. Intracellular expression of this antibody into either a stable cell line replicating subgenomic RNA, or a transient full-length HCV replication model, reduced both HCV RNA and viral protein expression. These results support the use of recombinant antibody fragments to inhibit NS3 enzyme as a novel, feasible, and effective approach for inhibiting HCV replication.






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Copyright © 2004 by the American Society for Investigative Pathology.