Centrifugal enhancement of hepatitis C virus infection of human hepatocytes.
Hepatitis C virus (HCV) is a human pathogen associated with chronic liver disease. Recently, the cell culture systems supporting complete replication and production of HCV genotype 2a (JFH1) have been established. This study investigated the effect of low-speed centrifugation on HCV JFH1 infection of human hepatocytes (Huh7.5.1). Higher levels of HCV RNA expression were observed in Huh7.5.1 cells infected with centrifugal inoculation of HCV JFH1 than those in the control cells. This increased HCV RNA expression was associated with the elevated expression of HCV NS3 protein in the hepatocytes. The centrifugal enhancement of HCV infection was time and speed dependent. However, the enhancement was not observed when centrifugation was performed before or after HCV infection. In addition, there was no association between centrifugal enhancement and the expression of HCV entry receptors (CD81 and claudin-1) and intracellular IFN-alpha in the hepatocytes. These data indicate that centrifugal inoculation is a useful tool for increasing the efficiency of HCV infection and replication in the target cells in vitro.
Ye
Li
L
Division of Allergy & Immunology, Joseph Stokes, Jr. Research Institute at The Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Wang
Xu
X
Wang
Shihong
S
Luo
Guangxiang
G
Wang
Yanjian
Y
Liang
Hao
H
Ho
Wenzhe
W
D015703:Antigens, CD / Q000096:biosynthesis; D002460:Cell Line; D002498:Centrifugation; D062445:Claudin-1; D015870:Gene Expression; D016174:Hepacivirus / Q000254:growth & development*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis; D008565:Membrane Proteins / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D060225:Tetraspanin 28; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis; D014776:Virus Cultivation / Q000379:methods*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D015703:Antigens, CD; D062445:Claudin-1; D016898:Interferon-alpha; D008565:Membrane Proteins; D012367:RNA, Viral; D060225:Tetraspanin 28; D017361:Viral Nonstructural Proteins; C554798:CD81 protein, human; C568108:CLDN1 protein, human; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases