Intrahepatic hepatitis C virus RNA quantification in microdissected hepatocytes

• Published in: Journal of hepatology 2004-04, Vol.40 (4), p.682-688
• Main Authors: Vona, Giovanna, Tuveri, Rosella, Delpuech, Oona, Vallet, Anais, Canioni, Danielle, Ballardini, Giorgio, Baptiste Trabut, Jean, Le Bail, Brigitte, Nalpas, Bertrand, Carnot, Françoise, Pol, Stanislas, Brechot, Christian, Thiers, Valérie
• Format: Article
• Language: English
• Subjects: Adult; Aged; Base Sequence; Biological and medical sciences; Dissection; DNA, Viral; DNA, Viral - genetics; Female; Gastroenterology. Liver. Pancreas. Abdomen; Hepacivirus; Hepacivirus - genetics; Hepacivirus - isolation & purification; Hepacivirus - physiology; Hepatitis C Antibodies; Hepatitis C virus; Hepatitis C, Chronic; Hepatitis C, Chronic - virology; Hepatocytes; Hepatocytes - virology; Human viral diseases; Humans; Infectious diseases; Intrahepatic HCV; Laser microdissection; Life Sciences; Male; Medical sciences; Middle Aged; RNA, Viral; RNA, Viral - analysis; RNA, Viral - blood; RNA, Viral - genetics; Viral diseases; Viral hepatitis; Viremia; Viremia - virology; Virus Replication
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/j.jhep.2003.12.012

Background/Aims: Debate continues on whether serum and intrahepatic HCV viral loads are correlated and if HCV viral load correlates with the severity of liver disease. These difficulties may at least in part be linked to liver cell heterogeneity, when total liver extracts from HCV-infected individuals are tested for HCV RNA quantification. We have therefore investigated the feasibility of quantifying HCV replication using a laser-based microdissection technique. Methods: We compared the results with those obtained for serum HCV RNA quantification and immunochemistry in the case of HCV antigen detection in the liver. Twenty-one HCV-positive patients with chronic active hepatitis ( n=10) or cirrhosis ( n=11) were analyzed. Results: A positive correlation ( P=0.0019) was observed between HCV RNA quantifications in sera and microdissected cells. Immunohistochemistry demonstrated that HCV antigen hepatocytes were randomly distributed within liver lobules. Their percentage varied in different patients (0–40%), but did not correlate with the HCV viral load. Conclusions: We have designed a sensitive methodology to evaluate the intrahepatic HCV viral load by combining a standardized RNA quantification method with microdissected hepatocytes from frozen liver needle biopsies. Our results directly demonstrate a positive correlation between serum and intrahepatic viral loads, which therefore provides a reliable reflection of intrahepatic HCV replication.