title: Monocyte-derived dendritic cells from HCV-infected patients transduced with an adenovirus expressing NS3 are functional when stimulated with the TLR3 ligand poly(I:C).
abstract: Dendritic cells (DC) transfected with an adenovirus encoding hepatitis C virus (HCV) NS3 protein (AdNS3) induce potent antiviral immune responses when used to immunize mice. However, in HCV infected patients, controversial results have been reported regarding the functional properties of monocyte-derived DC (MoDC), a cell population commonly used in DC vaccination protocols. Thus, with the aim of future vaccination studies we decided to characterize MoDC from HCV patients transfected with AdNS3 and stimulated with the TLR3 ligand poly(I:C). Phenotypic and functional properties of these cells were compared with those from MoDC obtained from uninfected individuals. PCR analysis showed that HCV RNA was negative in MoDC from patients after the culture period. Also, phenotypic analysis of these cells showed lower expression of CD80, CD86, and CD40, but similar expression of HLA-DR molecules as compared to MoDC from uninfected individuals. Functional assays of MoDC obtained from patients and controls showed a similar ability to activate allogeneic lymphocytes or to produce IL-12 and IL-10, although lower IFN-alpha levels were produced by cells from HCV patients after poly(I:C) stimulation. Moreover, both groups of MoDC induced similar profiles of IFN-gamma and IL-5 after stimulation of allogeneic T-cells. Finally, migration assays did not reveal any difference in their ability to respond to CCL21 chemokine. In conclusion, MoDC from HCV patients are functional after transduction with AdNS3 and stimulation with poly(I:C). These findings suggest that these cells may be useful for therapeutic vaccination in chronic HCV infection.
authors.affiliation: Division of Hepatology and Gene Therapy, Center for Applied Medical Research (CIMA), Pamplona, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Echeverría
authors.forename: A
authors.initials: A
authors.lastname: Zabaleta
authors.forename: L
authors.initials: L
authors.lastname: Silva
authors.forename: N
authors.initials: N
authors.lastname: Díaz-Valdés
authors.forename: J I
authors.initials: JI
authors.lastname: Riezu-Boj
authors.forename: J J
authors.initials: JJ
authors.lastname: Lasarte
authors.forename: F
authors.initials: F
authors.lastname: Borrás-Cuesta
authors.forename: M P
authors.initials: MP
authors.lastname: Civeira
authors.forename: J
authors.initials: J
authors.lastname: Prieto
authors.forename: P
authors.initials: P
authors.lastname: Sarobe
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000328:Adult; D000368:Aged; D015703:Antigens, CD / Q000032:analysis; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000737:chemistry / Q000276:immunology*; D005260:Female; D005822:Genetic Vectors; D006684:HLA-DR Antigens / Q000032:analysis; D016174:Hepacivirus / Q000276:immunology*; D006526:Hepatitis C / Q000276:immunology*; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D008297:Male; D008875:Middle Aged; D011070:Poly I-C / Q000494:pharmacology*; D051196:Toll-Like Receptor 3 / Q000819:agonists*; D014161:Transduction, Genetic; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D015703:Antigens, CD; D016207:Cytokines; D006684:HLA-DR Antigens; D007155:Immunologic Factors; D011070:Poly I-C; D051196:Toll-Like Receptor 3; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C495344:TLR3 protein, human