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.
Auffermann-GretzingerS.,
KeeffeE. B.,
LevyS.2001; Impaired dendritic cell maturation in patients with chronic, but not resolved, hepatitis C virus infection. Blood 97:3171–3176
BainC.,
FatmiA.,
ZoulimF.,
ZarskiJ. P.,
TrepoC.,
InchauspeG.2001; Impaired allostimulatory function of dendritic cells in chronic hepatitis C infection. Gastroenterology 120:512–524
BeldM.,
PenningM.,
LukashovV.,
McMorrowM.,
RoosM.,
PakkerN.,
van den HoekA.,
GoudsmitJ.1998; Evidence that both HIV and HIV-induced immunodeficiency enhance HCV replication among HCV seroconverters. Virology 244:504–512
BronowickiJ. P.,
LoriotM. A.,
ThiersV.,
GrignonY.,
ZignegoA. L.,
BrechotC.1998; Hepatitis C virus persistence in human hematopoietic cells injected into SCID mice. Hepatology 28:211–218
ChooQ. L.,
RichmanK. H.,
HanJ. H.,
BergerK.,
LeeC.,
DongC.,
GallegosC.,
CoitD.,
Medina-SelbyR.,
BarrP. J.,
WeinerA. J.,
BradleyD. W.,
KuoG.,
HoughtonM.1991; Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A 88:2451–2455
CookE. B.,
StahlJ. L.,
LoweL.,
ChenR.,
MorganE.,
WilsonJ.,
VarroR.,
ChanA.,
GrazianoF. M.,
BarneyN. P.2001; Simultaneous measurement of six cytokines in a single sample of human tears using microparticle-based flow cytometry: allergics vs. non-allergics. J Immunol Methods 254:109–118
DaarE. S.,
LynnH.,
DonfieldS.,
GompertsE.,
O'BrienS. J.,
HilgartnerM. W.,
HootsW. K.,
ChernoffD.,
ArkinS.,
WongW. Y.,
WinklerC. A.2001; Hepatitis C virus load is associated with human immunodeficiency virus type 1 disease progression in hemophiliacs. J Infect Dis 183:589–595
HilfenhausJ.,
GronerA.,
NowakT.,
WeimerT.1997; Analysis of human plasma products: polymerase chain reaction does not discriminate between live and inactivated viruses. Transfusion 37:935–940
LaneB. R.,
LoreK.,
BockP. J.,
AnderssonJ.,
CoffeyM. J.,
StrieterR. M.,
MarkovitzD. M.2001; Interleukin-8 stimulates human immunodeficiency virus type 1 replication and is a potential new target for antiretroviral therapy. J Virol 75:8195–8202
LanfordR. E.,
SureauC.,
JacobJ. R.,
WhiteR.,
FuerstT. R.1994; Demonstration of in vitro infection of chimpanzee hepatocytes with hepatitis C virus using strand-specific RT/PCR. Virology 202:606–614
LaskusT.,
RadkowskiM.,
WangL. F.,
VargasH.,
RakelaJ.1997; Lack of evidence for hepatitis G virus replication in the livers of patients coinfected with hepatitis C and G viruses. J Virol 71:7804–7806
LaskusT.,
RadkowskiM.,
PiasekA.,
NowickiM.,
HorbanA.,
CianciaraJ.,
RakelaJ.2000a; Hepatitis C virus in lymphoid cells of patients coinfected with human immunodeficiency virus type 1: evidence of active replication in monocytes/macrophages and lymphocytes. J Infect Dis 181:442–448
LaskusT.,
RadkowskiM.,
WangL. F.,
NowickiM.,
RakelaJ.2000b; Uneven distribution of hepatitis C virus quasispecies in tissues from subjects with end-stage liver disease: confounding effect of viral adsorption and mounting evidence for the presence of low-level extrahepatic replication. J Virol 74:1014–1017
LaskusT.,
WangL. F.,
RadkowskiM.,
VargasH.,
NowickiM.,
WilkinsonJ.,
RakelaJ.2001; Exposure of hepatitis C virus (HCV) RNA-positive recipients to HCV RNA-positive blood donors results in rapid predominance of a single donor strain and exclusion and/or suppression of the recipient strain. J Virol 75:2059–2066
LehmannC.,
SprengerH.,
NainM.,
BacherM.,
GemsaD.1996; Infection of macrophages by influenza A virus: characteristics of tumour necrosis factor-alpha (TNF alpha) gene expression. Res Virol 147:123–130
LeratH.,
RuminS.,
HabersetzerF.,
BerbyF.,
TrabaudM. A.,
TrepoC.,
InchauspeG.1998; In vivo tropism of hepatitis C virus genomic sequences in hematopoietic cells: influence of viral load, viral genotype, and cell phenotype. Blood 91:3841–3849
MaheY.,
MukaidaN.,
KunoK.,
AkiyamaM.,
IkedaN.,
MatsushimaK.,
MurakamiS.1991; Hepatitis B virus X protein transactivates human interleukin-8 gene through acting on nuclear factor κ B and CCAAT/enhancer-binding protein-like cis-elements. J Biol Chem 266:13759–13763
MoriN.,
MukaidaN.,
BallardD. W.,
MatsushimaK.,
YamamotoN.1998; Human T-cell leukemia virus type I Tax transactivates human interleukin 8 gene through acting concurrently on AP-1 and nuclear factor-kappa B-like sites. Cancer Res 58:3993–4000
MurayamaT.,
KunoK.,
JisakiF.,
ObuchiM.,
SakamuroD.,
FurukawaT.,
MukaidaN.,
MatsushimaK.1994; Enhancement human cytomegalovirus replication in a human lung fibroblast cell line by interleukin-8. J Virol 68:7582–7585
MurayamaT.,
OharaY.,
ObuchiM.,
KhabarK. S.,
HigashiH.,
MukaidaN.,
MatsushimaK.1997; Human cytomegalovirus induces interleukin-8 production by a human monocytic cell line, THP-1, through acting concurrently on AP-1- and NF- κ B-binding sites of the interleukin-8 gene. J Virol 71:5692–5695
NavasS.,
MartinJ.,
QuirogaJ. A.,
CastilloI.,
CarrenoV.1998; Genetic diversity and tissue compartmentalization of the hepatitis C virus genome in blood mononuclear cells, liver, and serum from chronic hepatitis C patients. J Virol 72:1640–1646
NowakT.,
NiedrigM.,
BernhardtD.,
HilfenhausJ.1993; Inactivation of HIV, HBV, HCV related viruses and other viruses in human plasma derivatives by pasteurisation. Dev Biol Stand 81:169–176
PolyakS. J.,
KhabarK. S.,
PaschalD. M.,
EzelleH. J.,
DuverlieG.,
BarberG. N.,
LevyD. E.,
MukaidaN.,
GretchD. R.2001a; Hepatitis C virus nonstructural 5A protein induces interleukin-8, leading to partial inhibition of the interferon-induced antiviral response. J Virol 75:6095–6106
PolyakS. J.,
KhabarK. S.,
RezeiqM.,
GretchD. R.2001b; Elevated levels of interleukin-8 in serum are associated with hepatitis C virus infection and resistance to interferon therapy. J Virol 75:6209–6211
SansonnoD.,
IacobelliA. R.,
CornacchiuloV.,
IodiceG.,
DammaccoF.1996; Detection of hepatitis C virus (HCV) proteins by immunofluorescence and HCV RNA genomic sequences by non-isotopic in situ hybridization in bone marrow and peripheral blood mononuclear cells of chronically HCV-infected patients. Clin Exp Immunol 103:414–421
ShimizuY. K.,
PurcellR. H.,
YoshikuraH.1993; Correlation between the infectivity of hepatitis C virus in vivo and its infectivity in vitro. Proc Natl Acad Sci U S A 90:6037–6041
ShimizuY. K.,
IgarashiH.,
KanematuT.,
FujiwaraK.,
WongD. C.,
PurcellR. H.,
YoshikuraH.1997; Sequence analysis of the hepatitis C virus genome recovered from serum, liver, and peripheral blood mononuclear cells of infected chimpanzees. J Virol 71:5769–5773
SimmondsP.,
HolmesE. C.,
ChaT. A.,
ChanS. W.,
McOmishF.,
IrvineB.,
BeallE.,
YapP. L.,
KolbergJ.,
UrdeaM. S.1993; Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. J Gen Virol 74:2391–2399
ThomasD. L.,
RichJ. D.,
SchumanP.,
SmithD. K.,
AstemborskiJ. A.,
NoltK. R.,
KleinR. S.2001; Multicenter evaluation of hepatitis C RNA levels among female injection drug users. J Infect Dis 183:973–976