Original Articles
Copyright ©The Author(s) 2001. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 15, 2001; 7(2): 228-234
Published online Apr 15, 2001. doi: 10.3748/wjg.v7.i2.228
HCV-RNA positivity in peripheral blood mononuclear cells of patients with chronic HCV infection: does it really mean viral replication?
Volker Meier, Sabine Mihm, Perdita Wietzke Braun, Guliano Ramadori
Volker Meier, Sabine Mihm, Perdita Wietzke Braun, Guliano Ramadori, Georg-August-Universität göttingen, Zentrum innere medizin, Abteilung FÜr Gastroenterologie Und Endokrinologie, Robert-Koch-Straβe 40, 37075 Göttingen, Germany
Author contributions: All authors contributed equally to the work.
Supported by a grant of DFG (SFB 402 Teilprojekt C1 (Mihm)) and by a grant of Hoffmann La Roche (Grenzach-Wyhden, Germany). Part of the data has been presented as poster at the 1999 EASL-meeting in Neaples.
Correspondence to: G. Ramadori, M.D., Georg-August-Universität Göttingen, Zentrum innere medizin, Abteilung FÜr Gastroenterologie Und Endokrinologie, Robert-Koch-Straβe 40, 37075 Göttingen, Germany. gramado@med.uni-goettingen.de
Telephone: +49-0551-396301 Fax: 0049-0551-398596
Received: February 6, 2001
Revised: February 22, 2001
Accepted: March 1, 2001
Published online: April 15, 2001
Abstract

AIM: To analyze the association of HCV-RNA with peripheral blood mononuclear cells (PBMC) and to answer the question whether HCV-RNA positivity in PBMC is due to viral replication.

METHODS: HCV-RNA was monitored in serum and PBMC preparations from 15 patients with chronic HCV infection before, during and after an IFN-α therapy using a nested RT/PCR technique. In a second approach, PBMC from healthy donors were incubated in HCV positive plasma.

RESULTS: In the IFN-α responding patients, HCV-RNA disappeared first from total RNA preparations of PBMC and then from serum. In contrast, in relapsing patients, HCV-RNA reappeared first in serum and then in PBMC. A quantitative analysis of the HCV-RNA concentration in serum was performed before and after transition from detectable to non detectable HCV-RNA in PBMC-RNA and vice versa. When HCV-RNA was detectable in PBMC preparations, the HCV concentration in serum was significantly higher than the serum HCV-RNA concentration when HCV-RNA in PBMC was not detectable. Furthermore, at no time during the observation period was HCV specific RNA observed in PBMC, if HCV-RNA in serum was under the detection limit. Incubation of PBMC from healthy donors with several dilutions of HCV positive plasma for two hours showed a concentration dependent PCR positivity for HCV-RNA in reisolated PBMC.

CONCLUSION: The detectability of HCV-RNA in total RNA from PBMC seems to depend on the HCV concentration in serum. Contamination or passive adsorption by circulating virus could be the reason for detection of HCV-RNA in PBMC preparations of chronically infected patients.

Keywords: hepatitis C like viruses; hepatitis C, chronic; RNA, viral/blood; virus replication; monocytes; interferon alpha/therapeutic use; polymerase chain reaction