Intrahepatic hepatitis C virus RNA quantification in microdissected hepatocytes.
OBJECTIVE
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.
Vona
Giovanna
G
INSERM U370-Pasteur, Necker Faculty, Paris, France.
Tuveri
Rosella
R
Delpuech
Oona
O
Vallet
Anais
A
Canioni
Danielle
D
Ballardini
Giorgio
G
Baptiste Trabut
Jean
J
Le Bail
Brigitte
B
Nalpas
Bertrand
B
Carnot
Françoise
F
Pol
Stanislas
S
Brechot
Christian
C
Thiers
Valérie
V
D000328:Adult; D000368:Aged; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004210:Dissection; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D018937:Hepatitis C Antibodies; D019698:Hepatitis C, Chronic / Q000821:virology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood / Q000235:genetics; D014766:Viremia / Q000821:virology; D014779:Virus Replication
D004279:DNA, Viral; D018937:Hepatitis C Antibodies; D012367:RNA, Viral