Characterization of monoclonal antibodies against the nonstructural 5A protein of hepatitis C virus.
The nonstructural 5A (NS5A) protein of hepatitis C virus (HCV) is a multifunctional protein that leads to pleiotropic responses, in part by regulating cell growth and cellular signaling pathways. Here, we produced monoclonal antibodies (mAbs) directed against the HCV NS5A protein. The N-terminal epitope was mapped to amino acids 60-80 of the NS5A protein, and the epitope in the middle region was mapped to amino acids 221-236. Because these epitopes overlap with binding regions of human vesicle-associated membrane-protein-associated protein (hVAP)-B and TNF-receptor-associated factor 2 (TRAF2), respectively, we investigated these mAbs for their potential capacity to inhibit viral and cellular interactions. We found that NS5A and hVAP-B interaction was abolished by mAb E5D3, and NS5A and TRAF2 interaction was inhibited by mAb C6D4. Since hVAP-B is necessary for HCV replication and TRAF2 is the major transducer in TNF signaling cascades, these data may provide further insights into the mechanisms underlying HCV replication and viral modulation of host signal transduction.
Kang
Sang Min
SM
National Research Laboratory of Hepatitis C Virus, Ilsong Institute of Life Science, Hallym University, 1605-4 Gwanyang-dong, Dongan-gu, Anyang 431-060, Korea.
Jun
Hyun-Jung
HJ
Lim
Yun-Sook
YS
Choi
Soo-Ho
SH
Hwang
Soon B
SB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002460:Cell Line; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005260:Female; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D047992:TNF Receptor-Associated Factor 2 / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
D000911:Antibodies, Monoclonal; D000939:Epitopes; D047992:TNF Receptor-Associated Factor 2; D033921:Vesicular Transport Proteins; D017361:Viral Nonstructural Proteins; C496748:VAPB protein, human; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase