Cell Attachment and Penetration by Influenza Virus

Abstract

Attachment and penetration of influenza virus into clone 1-5C-4 cells were quantitatively determined by the immunofluorescent cell-counting assay. Aided by centrifugal force, more than 95% of virus inocula of five representative influenza virus strains (A 0 /PR8, A 1 /Ann Arbor, A 2 /Japan, B/Lee, B/Great Lakes) were attached to cells at a linear rate within 10 min, in contrast to approximately 35% after stationary incubation at 35 C for 2 h. By the former procedure, a proportionality between the number of infected cells and volume of inoculum was revealed which was not evident when stationary incubation was employed. Maximal binding of virus to cells occurred at 0.2 M NaCl. The salt requirement, added to evidence of pH dependence and temperature independence, indicated that the initial virus-cell union involved electrostatic forces. Virus penetration into cells, measured by the insensitivity of virus-cell complexes to antiviral serum, was linear and complete within 15 min at 35 C for all five virus strains tested. Maximal virus penetration occurred at 0.1 to 0.2 M NaCl; the process was pH- and temperature-dependent. Both virus attachment and penetration processes were partially inhibited in the presence of diethylaminoethyl-dextran.