title: Presence of hepatitis C virus (HCV)-RNA in peripheral blood mononuclear cells in HCV serum negative patients during interferon and ribavirin therapy.
abstract: Identification of hepatitis C virus (HCV)-RNA in blood serum is crucial for hepatitis C diagnosis and for appropriate treatment. Detection of HCV-RNA in blood serum is used for therapy monitoring of patients with hepatitis C. Despite HCV-RNA elimination from blood serum during treatment in some patients, HCV viremia appears again after the completion of therapy. The aim of this study was to assess HCV-RNA in peripheral blood mononuclear cells (PBMCs) of hepatitis C patients in relation to HCV-RNA and antibodies to HCV in the serum. The study involved 71 patients undergoing anti-viral therapy (interferon and ribavirin). RNA isolated from serum and PBMCs was examined for the presence of HCV-RNA by an RT-PCR technique using specific oligonucleotide primers or by commercially available kits. In order to show the possible presence of HCV sequences in PBMCs, molecular DNA probes were constructed with a PCR amplicon and biotin-labelled by nick translation, and FISH and extended chromatin fibers in situ hybridization (ECFs-FISH) techniques were used. A 24-month follow-up study revealed that 34 out of 59 patients (58%) eliminated HCV-RNA from their sera. In the serum negative group, HCV-RNA was detected in PBMCs of 2 patients. The presence of HCV-RNA in PBMCs was confirmed by the FISH technique. In the ECFs-FISH procedure, no signal was found in all examined patients. Our data suggest that PBMCs infected with HCV can serve as a virus reservoir. HCV-RNA serum negative patients who have HCV-RNA in their leukocytes after completion of anti-viral therapy would be at great risk of hepatitis C recurrence. These HCV-RNA serum negative but PBMCs positive patients would be a potential source of HCV spread.
authors.affiliation: University of Medical Sciences, Poznan, Poland.
authors.forename: Danuta
authors.initials: D
authors.lastname: Januszkiewicz-Lewandowska
authors.forename: Jacek
authors.initials: J
authors.lastname: Wysocki
authors.forename: Monika
authors.initials: M
authors.lastname: Pernak
authors.forename: Karina
authors.initials: K
authors.lastname: Nowicka
authors.forename: Mariola
authors.initials: M
authors.lastname: Zawada
authors.forename: Jolanta
authors.initials: J
authors.lastname: Rembowska
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Lewandowski
authors.forename: Przemysław
authors.initials: P
authors.lastname: Mańkowski
authors.forename: Jerzy
authors.initials: J
authors.lastname: Nowak
mesh_terms: D000293:Adolescent; D000914:Antibodies, Viral / Q000097:blood; D000998:Antiviral Agents / Q000627:therapeutic use; D002648:Child; D002675:Child, Preschool; D004359:Drug Therapy, Combination; D005260:Female; D016174:Hepacivirus / Q000235:genetics*; D019698:Hepatitis C, Chronic / Q000097:blood* / Q000188:drug therapy* / Q000821:virology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007372:Interferons / Q000627:therapeutic use*; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000097:blood*; D012254:Ribavirin / Q000627:therapeutic use*
chemical_list: D000914:Antibodies, Viral; D000998:Antiviral Agents; D012367:RNA, Viral; D012254:Ribavirin; D007372:Interferons