TY - JOUR
T1 - Infection of primary human macrophages with hepatitis C virus in vitro
T2 - Induction of tumour necrosis factor-α and interleukin 8
AU - Radkowski, Marek
AU - Bednarska, Agnieszka
AU - Horban, Andrzej
AU - Stanczak, Janusz
AU - Wilkinson, Jeffrey
AU - Adair, Debra M.
AU - Nowicki, Marek
AU - Rakela, Jorge
AU - Laskus, Tomasz
PY - 2004/1
Y1 - 2004/1
N2 - 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-α (TNF-α) and interleukin (IL)-1β, 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-α 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-α and IL-8.
AB - 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-α (TNF-α) and interleukin (IL)-1β, 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-α 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-α and IL-8.
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U2 - 10.1099/vir.0.19491-0
DO - 10.1099/vir.0.19491-0
M3 - Article
C2 - 14718619
AN - SCOPUS:1642544601
SN - 0022-1317
VL - 85
SP - 47
EP - 59
JO - Journal of General Virology
JF - Journal of General Virology
IS - 1
ER -