Improved attachment of natural HCV isolate to Daudi cells upon elimination of immune complexes and close pH control.
OBJECTIVE
To study the viral requirements for attachment, entry and infection using natural isolates of HCV (infectious plasma of a patient infected with HCV from genotype 1b) and a cell line which has previously been shown to support HCV replication (Daudi cells).


METHODS
We studied attachment of HCV to Daudi cells, a human B-cell line. Quantification was done by real-time RT-PCR.


RESULTS
The best attachment levels were obtained using plasma depleted of immunocomplexes. Results of kinetics of HCV attachment to Daudi cells show that attachment is maximum at pH 7.0, and that it decreased drastically at any other pH studied (5.5, 6.0, 6.5, 7.5 and 8.0).


CONCLUSIONS
Depletion of immunocomplexes and pH control during infection are important parameters to improve attachment of HCV to Daudi cells using plasma as a natural source of virus.
Quer
Josep
J
Liver Unit, Department of Medicine, Hospital Universitari Vall d'Hebron, Barcelona, Spain. jquer@vhebron.net
Cos
Joan
J
Murillo
Pedro
P
Esteban
Juan Ignacio
JI
Esteban
Rafael
R
Guardia
Jaume
J
D000936:Antigen-Antibody Complex; D001402:B-Lymphocytes / Q000821:virology*; D002460:Cell Line; D019286:Endopeptidase K / Q000378:metabolism; D016174:Hepacivirus / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010949:Plasma / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014776:Virus Cultivation / Q000379:methods*
D000936:Antigen-Antibody Complex; D012367:RNA, Viral; D019286:Endopeptidase K