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Quantification of hepatitis C virus-infected peripheral blood mononuclear
cells by in situ reverse transcriptase-polymerase chain reaction
L Muratori, D Gibellini, M Lenzi, M Cataleta, P Muratori, MC Morelli and FB Bianchi
Cattedra di Medicina Interna II, Universita di Bologna, Policlinico S.
Orsola, Italy.
Hepatitis C virus (HCV) is known to infect peripheral blood mononuclear
cells (PBMC) of patients with chronic hepatitis C, but the proportion of
HCV-infected circulating cells is not detectable by conventional reverse
transcriptase-polymerase chain reaction (RT-PCR) and the pathogenic
significance of HCV lymphotropism is still unclear. Therefore, we have
devised an in situ RT-PCR technique using fluorescein-labeled HCV-specific
primers revealed by flow cytometry. PBMC were isolated from 28 patients
with chronic HCV-related liver disease; of these, 6 had previously received
an orthotopic liver transplantation (OLT) and were on immuno-suppressive
treatment. Fourteen patients (50%) were found positive for HCV genome
within PBMC by in situ RT-PCR, the proportion of HCV-infected cells ranging
from 0.2% to 8.1%. All 6 OLT patients tested positive. The fluorescent
signal, corresponding to the HCV-specific 340-bp amplicon, was confined to
part of the cytoplasmic compartment of scattered PBMC. Of these 14
patients, 12 had also negativestrand HCV RNA within PBMC detected by
"tagged" RT-PCR. We conclude that HCV may infect a significant proportion
of PBMC in chronic hepatitis C patients, especially immunosuppressed OLT
cases, and that viral replication within PBMC is a common occurrence. Over
time, the persistence of HCV-infected immune system cells might interfere
with normal immunologic mechanisms and play a role in the pathogenic
processes leading to extrahepatic disorders such as mixed cryoglobulinemia
and B-cell malignant lymphoma.
Volume 88,
Issue 7,
pp. 2768-2774,
10/01/1996
Copyright © 1996 by The American Society of Hematology
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