Buoyant density of hepatitis C virus recovered from infected hosts: two different features in sucrose equilibrium density-gradient centrifugation related to degree of liver inflammation.
Hepatitis C virus is reported to have a low buoyant density in sucrose. To determine the density of hepatitis C virus in the circulation of infected hosts and its association with the degree of liver inflammation, we examined serum samples from 10 patients who were positive for both hepatitis C virus antibody (C100 antigen) antibody and serum hepatitis C virus RNA. After the serum was ultracentrifuged in sucrose density gradient (10% to 60%), the hepatitis C virus RNA titer in each collected fraction was quantified by means of competitive reverse transcription-polymerase chain reaction. In samples from five blood donors, the hepatitis C virus RNA titer had a single peak at fractions with densities of 1.08 to 1.11 gm/ml. In samples from five patients with ALT abnormalities, the titer had two peaks at fractions with 1.09 to 1.10 gm/ml and 1.22 to 1.25 gm/ml. After the selected samples were treated with detergents and ultracentrifuged, the titer in the 1.08 to 1.11 gm/ml fractions decreased and that in the 1.22 to 1.25 gm fractions increased. This result implied that the hepatitis C virus density changed with removal of the viral envelope by lipid solvents. Thus the buoyant density of hepatitis C virus in sucrose was 1.08 to 1.11 gm/ml for an intact virion and 1.22 to 1.25 gm/ml for what was presumed to be a nucleocapsid. These results demonstrated that HCV virion is a dominant form in the circulation of blood donors without ALT abnormalities. In patients with liver inflammation HCV particles with higher densities of 1.22 to 1.25 gm/ml coexist with virion in the circulation, which might be presumed nucleocapsids.
Kanto
T
T
First Department of Medicine, Osaka University Medical School, Japan.
Hayashi
N
N
Takehara
T
T
Hagiwara
H
H
Mita
E
E
Naito
M
M
Kasahara
A
A
Fusamoto
H
H
Kamada
T
T
D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D002499:Centrifugation, Density Gradient; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006526:Hepatitis C / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D008099:Liver / Q000473:pathology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood; D014158:Transcription, Genetic
D017931:DNA Primers; D012367:RNA, Viral