title: Molecular and cellular determinants of cell-to-cell transmission of HCV in vitro.
abstract: It was reported previously that HCV can be transmitted from persistently infected human bone-marrow-derived B-lymphoblastoid cells (TO.FE(HCV)) to human hepatoma cells by cell-to-cell contact. The present study confirms and characterize further such type of HCV infection in vitro. TO.FE(HCV) cells were co-cultured with 2.2.15 hepatoma cells, that are not susceptible to cell-free infection by sera containing HCV of 1b genotype. By this co-cultivation system it was demonstrated that HCV transmission to recipient cells requires de novo virus RNA replication. Several factors may favor HCV-transmission, evidence is provided that TO.FE(HCV) cells were able to select HCV-quasispecies. 5'-UTR and core sequence analysis revealed differences in the HCV-quasispecies composition in serum inoculum and in infected TO.FE B-cells at 4 months post-inoculation. It is considered that the latter may be more successful in replicating HCV in vitro and used to express surface molecules which may be involved in cell-to-cell contact. In TO.FE(HCV) cells replicate distinct, or few close related, HCV-variants correlated with those of serum inoculum. Comparative analysis of tetra-spans and integrins expression undertaken by cytofluorimetry displayed higher level of expression for TO.FE cells in comparison to other human bone-marrow-derived B-cell lines. Overall, the observed persistent in vitro HCV replication is mediated by a continuous cell-to-cell reinfection that may be favored by selection of viral variants and expression of molecules involved in cell adhesion. These observations may provide an explanation for the establishment of HCV infection, the occurrence of chronic infection and HCV-related lymphoproliferative diseases.
authors: Institute of Neurobiology and Molecular Medicine, National Research Council (CNR), Roma, Italy.
authors: Maria Beatrice
authors: MB
authors: Valli
authors: Annalisa
authors: A
authors: Crema
authors: Giulia
authors: G
authors: Lanzilli
authors: Annalucia
authors: A
authors: Serafino
authors: Luisa
authors: L
authors: Bertolini
authors: Gianpietro
authors: G
authors: Ravagnan
authors: Antonio
authors: A
authors: Ponzetto
authors: Stefano
authors: S
authors: Menzo
authors: Massimo
authors: M
authors: Clementi
authors: Guido
authors: G
authors: Carloni
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D001402:B-Lymphocytes / Q000378:metabolism / Q000821:virology; D001483:Base Sequence; D002450:Cell Communication; D002460:Cell Line; D018920:Coculture Techniques; D005434:Flow Cytometry; D016174:Hepacivirus / Q000502:physiology*; D006526:Hepatitis C / Q000378:metabolism / Q000821:virology*; D022781:Hepatocytes; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics*; D017062:Small-Area Analysis; D014758:Viral Core Proteins / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D008565:Membrane Proteins; D012367:RNA, Viral; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus