Infection of primary human macrophages with hepatitis C virus in vitro: induction of tumour necrosis factor-alpha and interleukin 8.
Hepatitis C virus (HCV) has been reported to replicate in monocytes/macrophages in infected patients. However, it is unclear whether macrophages are susceptible to infection in vitro and whether such an infection is consequential. Sera from 26 HCV-infected patients were incubated with primary human macrophages collected from healthy donors. Virus negative strand was detected by a Tth enzyme-based strand-specific assay and virus sequences were analysed by single strand conformation polymorphism (SSCP) and sequencing. Concentrations of the cytokines tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta, IL-6, IL-8, IL-10 and IL-12p70 were measured in culture supernatants and respective mRNAs were analysed in cell extracts by quantitative RT-PCR. For 15 sera, HCV RNA was detectable in 2- and 3-week cultures from at least one donor. Virus negative strand was detected in 29 % of macrophage samples in this group. In four cases, HCV RNA sequences amplified from macrophages differed from those amplified from sera suggesting evolution during infection. Concentrations of TNF-alpha and IL-8 were found to be significantly higher in supernatants from HCV-infected cultures. In conclusion, these preliminary data suggest that primary human macrophages are susceptible to HCV infection in vitro and this infection is associated with the induction of cytokines TNF-alpha and IL-8.
Radkowski
Marek
M
Department of Medicine, SC Johnson Bldg Sj3, Mayo Clinic Scottsdale, AZ 85259, USA.|Institute of Infectious Diseases, Warsaw Medical Academy, Poland.
Bednarska
Agnieszka
A
Municipal Hospital for Infectious Diseases, Warsaw, Poland.
Horban
Andrzej
A
Municipal Hospital for Infectious Diseases, Warsaw, Poland.
Stanczak
Janusz
J
Municipal Hospital for Infectious Diseases, Warsaw, Poland.
Wilkinson
Jeffrey
J
Department of Medicine, SC Johnson Bldg Sj3, Mayo Clinic Scottsdale, AZ 85259, USA.
Adair
Debra M
DM
Department of Medicine, SC Johnson Bldg Sj3, Mayo Clinic Scottsdale, AZ 85259, USA.
Nowicki
Marek
M
Maternal-Child Virology Research Laboratory, University of Southern California, Los Angeles, CA 90033, USA.
Rakela
Jorge
J
Department of Medicine, SC Johnson Bldg Sj3, Mayo Clinic Scottsdale, AZ 85259, USA.
Laskus
Tomasz
T
Department of Medicine, SC Johnson Bldg Sj3, Mayo Clinic Scottsdale, AZ 85259, USA.
D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D016174:Hepacivirus / Q000276:immunology / Q000472:pathogenicity*; D006526:Hepatitis C / Q000276:immunology / Q000821:virology; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis*; D008264:Macrophages / Q000276:immunology / Q000821:virology*; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D017077:Culture Media, Conditioned; D016207:Cytokines; D016209:Interleukin-8; D012333:RNA, Messenger; D012367:RNA, Viral; D014409:Tumor Necrosis Factor-alpha