A study of susceptibility of primary human Kupffer cells to hepatitis C virus.
OBJECTIVE
Histological examination of a liver-biopsy from a patient with chronic hepatitis C shows activated Kupffer cells. In vitro infection of human Kupffer cells (KC) was performed to study their interaction with hepatitis C virus (HCV).


METHODS
KC, isolated by collagenase perfusion and centrifugal elutriation, were infected with various HCV positive sera. The presence of the viral genome was followed, at different times, quantitatively by a branched-DNA assay and qualitatively by reverse transcriptase-nested polymerase chain reaction. A strand-specific assay performed with the thermostable enzyme rTth was used to detect the synthesis of a negative replicative intermediate. Cytopathic effect was examined by electron microscopy. Production of cytokines and inducible nitric oxide synthase was evaluated in the supernatants.


RESULTS
Quantification of HCV-RNA showed that the level of viral RNA associated with KC after adsorption decreased rapidly. Genomic viral RNA disappeared within 5 days of infection. Negative-strand RNA was never detected in any of these experiments. No cytopathic effects could be detected at any time. KC did not produce inflammatory nor antiviral cytokines.


CONCLUSIONS
Our results strongly suggest that primary cultures of KC are not permissive for HCV in vitro.
Royer
Cathy
C
Institut de Virologie de la Faculté de Médecine de Strasbourg, INSERM U544, 3 Rue Koeberlé, 67000 Strasbourg, France.
Steffan
Anne-Marie
AM
Navas
Maria-Cristina
MC
Fuchs
Anne
A
Jaeck
Daniel
D
Stoll-Keller
Françoise
F
D004198:Disease Susceptibility; D016174:Hepacivirus / Q000235:genetics; D019698:Hepatitis C, Chronic / Q000473:pathology / Q000503:physiopathology* / Q000821:virology*; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D007728:Kupffer Cells / Q000378:metabolism / Q000821:virology*; D008854:Microscopy, Electron; D012367:RNA, Viral / Q000378:metabolism; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D016898:Interferon-alpha; D015850:Interleukin-6; D012367:RNA, Viral; D014409:Tumor Necrosis Factor-alpha