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Journal of Histochemistry and Cytochemistry, Vol. 46, 653-660, May 1998, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Detection and Localization by In Situ Molecular Biology Techniques and Immunohistochemistry of Hepatitis C Virus in Livers of Chronically Infected Patients

Francine M. Walkera, Marie-Christine Dazzaa,c, Marie-Christine Daugea,c, Olivier Bouchera, Christophe Bedela, Dominique Henina, and Therese Lehyb
a Department of Pathology, Hôpital Bichat-Claude Bernard, Paris, France
b INSERM U10, Hôpital Bichat-Claude Bernard, Paris, France
c INSERM U13, Hôpital Bichat-Claude Bernard, Paris, France

Correspondence to: Francine M. Walker, Service d’Anatomie Pathologique, Hôpital Bichat-Claude Bernard, 46 rue Henri Huchard, 75877 Paris Cédex 18, France.

Hepatitis C virus (HCV) detection in the livers of chronically infected patients remains a debatable issue. We used immunohistochemistry, in situ hybridization (ISH) alone or after microwave heating with FITC-labeled probes, RT-PCR with unlabeled primers followed by ISH (RT-PCR–ISH), and in situ RT-PCR with FITC-labeled primers (in situ RT-PCRd) to localize the virus in 38 liver biopsy specimens from 21 chronically infected HCV patients treated with interferon-{alpha} (IFN-{alpha}). Biopsies were taken at the beginning and end of IFN-{alpha} treatment and 1 year later. Results were compared with that of HCV-PCR in serum. RT-PCR–ISH and in situ RT-PCRd showed HCV signal in all liver biopsies even in responders with seronegative HCV PCR. This signal was intranuclear, diffuse, or peripheral, in hepatocytes, bile ductule cells, and lymphocytes. Cytoplasmic signals were occasionally observed. Whereas the percentage of labeled hepatocytes remained constant, the number of labeled lymphoid follicles decreased after INF-{alpha} therapy. Immunohistochemistry resulted in the same pattern of positivity but it was weaker and inconstant. This study indicates the persistency of HCV latency in IFN-{alpha} responders 1 year after IFN-{alpha} treatment cessation, a finding that certainly deserves confirmation. (J Histochem Cytochem 46:653–660, 1998)

Key Words: HCV, interferon-{alpha} therapy, in situ RT-PCR, in situ RT-PCRd, RT-PCR–ISH, immunohistochemistry, C100-3 protein, human liver


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