Evolution of viral quasispecies in interferon-treated patients with chronic hepatitis C virus infection.
OBJECTIVE
To evaluate whether interferon treatment failure/relapse is related to changes in hepatitis C virus quasispecies complexity (number of variants) or diversity (genetic relatedness of variants).


METHODS
We analyzed hypervariable region heterogeneity in hepatitis C virus-infected patients by heteroduplex mobility assay and by phylogenetic analysis of sequenced clones. Sera from 11 patients were tested. Response was defined biochemically and virologically. Patients were treated with 3 or 6 MIU interferon for 6 months and followed up for 6 months. Four patients were non-responders, four were transient responders and three untreated patients served as controls. Three time points were studied for the non-responders (pre-interferon, end of interferon, end of 6 months of follow-up), two for the transient responders (pre-interferon and post follow-up) and two for the controls (1 year apart). A total of 260 clones were examined by heteroduplex mobility assay and 144 clones were sequenced.


RESULTS
A linear correlation between heteroduplex mobility and nucleotide substitutions was observed, validating this method for assessment of quasispecies diversity. Although complexity at each time point was similar in all groups, diversity increased significantly with interferon treatment. The percentage of new variants in follow up was significantly higher in non-responders than in controls. These new variants exhibited a greater change in heteroduplex mobility, a higher percentage of changes in amino acids in non-responders compared to controls and were found to cluster separately from pretreatment variants when analyzed phylogenetically. These changes were less marked in transient responders.


CONCLUSIONS
These mutations may allow hepatitis C virus to escape antiviral effects of interferon therapy.
Hassoba
H M
HM
Department of Veterans Affairs Medical Center, University of California, San Francisco 94121, USA.
Bzowej
N
N
Berenguer
M
M
Kim
M
M
Zhou
S
S
Phung
Y
Y
Grant
R
R
Pessoa
M G
MG
Wright
T L
TL
D000595:Amino Acid Sequence / Q000235:genetics; D000998:Antiviral Agents / Q000627:therapeutic use*; D005075:Biological Evolution; D004351:Drug Resistance; D014644:Genetic Variation / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics*; D019698:Hepatitis C, Chronic / Q000188:drug therapy* / Q000821:virology*; D020180:Heteroduplex Analysis; D006801:Humans; D007372:Interferons / Q000627:therapeutic use*; D008969:Molecular Sequence Data; D012016:Reference Values
D000998:Antiviral Agents; D007372:Interferons