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Blood, Vol. 94 No. 4 (August 15), 1999:
pp. 1183-1191
Low Levels of Hepatitis C Virus RNA in Serum, Plasma, and Peripheral
Blood Mononuclear Cells of Injecting Drug Users During Long
Antibody-Undetectable Periods Before Seroconversion
Marcel Beld,
Maarten Penning,
Marieke van Putten,
Anneke van den
Hoek,
Marjolein Damen,
Michèl R. Klein, and
Jaap Goudsmit
From the Academic Medical Centre, University of Amsterdam, Department
of Human Retrovirology, Amsterdam, The Netherlands; the Municipal
Health Service, Department of Public Health and Environment, Amsterdam,
The Netherlands; the Central Laboratory of The Netherlands Red Cross
Blood Transfusion Service, Amsterdam, The Netherlands; and
the Medical Research Council (MRC) Laboratories, Banjul, The Gambia.
Screening of antibodies to hepatitis C virus (HCV) is widely used
for monitoring the prevalence of HCV infections and to assess HCV
infectivity. Among HCV-infected individuals in the general population,
the interval between the detection of HCV RNA and the development of
HCV antibodies is usually 5 to 6 weeks, but in rare cases,
seroconversion may be prolonged up to 6 to 9 months. In this study, we
tested for the presence of HCV RNA during the antibody-undetectable
period of 19 drug-injecting HCV seroconverters to gain insight into the
antibody-negative carrier status in this population. HCV seroconversion
status was determined by testing the first and last serum samples
obtained from each subject, using third-generation antibody screening
and confirmation assays. Serial samples were tested for HCV-specific
antibodies to establish the moment of seroconversion and HCV RNA by
single reverse transcriptase-polymerase chain reaction (RT-PCR) and
branched DNA assay (bDNA) in serum. Plasma and peripheral blood
mononuclear cells (PBMCs) were independently collected and tested for
HCV RNA. HCV RNA-positivity was confirmed by Southern blot
hybridization and sequencing of serial samples. The 19 HCV
seroconverters had a mean follow-up of 5 years (range, 1 to 8 years).
Of the 19, 4 were human immunodeficiency virus (HIV)-infected before
HCV seroconversion. HCV RNA was detected in serum before seroconversion
in 12 (63.2%) of the 19 HCV seroconverters, independent of HIV status.
In 7 of these 12, the antibody-undetectable period was relatively short
(2 to 10 months). The other 5, who were all HIV-negative before HCV
seroconversion, had intermittent low levels of HCV RNA before
seroconversion for a period of more than 12 months, with a mean of 40.8 months (range, 13 to 94 months). In all 5 individuals, independent
repeats of the experiments confirmed the presence of HCV RNA in serum,
and in 3 of these individuals, HCV-positivity was confirmed in
independently collected plasma and PBMC samples. Low levels of HCV RNA
may be present during prolonged antibody-undetectable periods before
seroconversion in a number of injecting drug users. Independent of HIV
status, their immune system appears to be unable to respond to these
low HCV RNA levels and was sometimes only activated after reinfections with distinct HCV genotypes. These results indicate that primary HCV
infection may not always elicit the rapid emergence of HCV antibodies
and suggests that persistent low levels of HCV RNA (regardless of the
genotype) may not elicit at all or delay antibody responses for
prolonged periods of time.
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