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Fig. 4. Construction of pseudotype retroviruses. 293T cells are trans-
fected with three plasmids encoding HCV E1 and E2 proteins, retroviral
gag-pol proteins, and a packaging competent GFP, respectively. After 2 to 3
days incubation, pseudotype retroviruses secreted into the culture super-
natants. Infectivity of pseudotype viruses containing Gag, Pol, E1 and E2
proteins, as well as the RNA encoding the GFP or luciferase gene is evalu-
ated by the expression of GFP or Luciferase in target cells.
three kinds of expression plasmids (Fig. 4) are infectious to Huh7 cells
but not to HepG2 cells. In addition, infection of the pseudotype parti-
cles was neutralized by anti-E2 monoclonal antibodies as well as sera of
most of patients infected with HCV. Therefore, pseudotype retroviruses
also represent a useful model for the study of the early steps of HCV life
cycle in combination with the pseudotype VSVs. It was also shown that
infection of the pseudotype retroviruses was required both E1 and E2
proteins and no significant difference on the infectivity and neutraliza-
tion properties was observed among HCV genotypes. 8,9,44,52 Although
pseudotype retrovirus infection was inhibited by the addition of anti-
hCD81 antibody or a soluble form of hCD81, expression of hCD81
alone failed to confer mouse NIH3T3 cells susceptible to infection by
pseudotype MLVs. 10 A similar study was demonstrated that pseudotype
HIVs possessing unmodified HCV envelope proteins infect Huh7 cells
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