Hepatitis C virus-RNAs in plasma and in peripheral blood mononuclear cells of hemophiliacs with chronic hepatitis C: evidence for viral replication in peripheral blood mononuclear cells.
Hemophiliacs who have been exposed to unheated and/or dry heated pooled clotting factor concentrates are at high risk of chronic hepatitis C. Although the mechanism and site of hepatitis C virus (HCV) replication are not yet known, HCV is thought to replicate through a complementary negative RNA strand, as has been shown for flaviviruses. The detection of negative RNA strands has therefore been regarded as a marker of replication. We investigated the prevalence of HCV-RNA and of negative HCV-RNA strands in peripheral blood mononuclear cells (PBMC) and plasma of hemophiliacs. Forty-three of 47 patients studied (91%) had anti-HCV antibodies and in 36 patients HCV-RNA was detectable in PBMC. In one group of 20 patients negative HCV-RNA strands were present in PBMC and 10 of these patients also had negative HCV-RNA strands in plasma. In another group of nine patients HCV-RNA was detected in PBMC, although cDNA synthesis was carried out in the absence of primers. Only in two of these nine patients negative and positive HCV-RNA strands were demonstrated specifically in PBMC using a modified reverse transcription step. If the presence of negative HCV-RNA strands can be considered as marker of viral replication, the findings indicate that HCV can replicate in PBMC. Furthermore, in certain patients it is impossible to use the currently available technique to detect selectively positive or negative HCV-RNA strands.
Willems
M
M
Division of Liver and Pancreatic Diseases, University Hospital Gasthuisberg, Leuven, Belgium.
Peerlinck
K
K
Moshage
H
H
Deleu
I
I
Van den Eynde
C
C
Vermylen
J
J
Yap
S H
SH
D000293:Adolescent; D000328:Adult; D000368:Aged; D000410:Alanine Transaminase / Q000097:blood; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D005260:Female; D015658:HIV Infections / Q000150:complications; D006467:Hemophilia A / Q000097:blood / Q000150:complications* / Q000382:microbiology; D002836:Hemophilia B / Q000097:blood / Q000150:complications / Q000382:microbiology; D016174:Hepacivirus / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D006508:Hepatitis Antibodies / Q000097:blood; D006509:Hepatitis B / Q000097:blood / Q000150:complications / Q000201:enzymology / Q000276:immunology; D006510:Hepatitis B Antibodies / Q000097:blood; D006526:Hepatitis C / Q000150:complications / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D018937:Hepatitis C Antibodies; D006521:Hepatitis, Chronic / Q000150:complications / Q000382:microbiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood*; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D065227:Transfusion Reaction; D014766:Viremia / Q000201:enzymology / Q000276:immunology / Q000382:microbiology*; D014779:Virus Replication
D006508:Hepatitis Antibodies; D006510:Hepatitis B Antibodies; D018937:Hepatitis C Antibodies; D012367:RNA, Viral; D000410:Alanine Transaminase; D012194:RNA-Directed DNA Polymerase