Attachment of infectious influenza A viruses of various subtypes to live mammalian and avian cells as measured by flow cytometry.
At present there is much interest in the cell tropism and host range of influenza viruses, especially those of the H5N1 subtype. We wished to develop a method that would enable investigation of attachment of infectious virus through the interaction of the hemagglutinin molecule and live mammalian and avian cells and the subsequent infection of these cells. To this end, influenza viruses of various HA subtypes were constructed that either carry the green fluorescent protein (GFP) instead of the neuraminidase protein, or that express GFP in the cytoplasm of infected cells. The HA genes were derived from influenza viruses A/PR/8/34 (H1N1), A/Netherlands/178/95 (H3N2) and A/Vietnam/1194/04 (H5N1). Using these pairs of viruses, attachment and post-attachment events in the virus replication cycle can be distinguished. In general, the expression of NeuAc(alpha2-3)Gal or NeuAc(alpha2-6)Gal receptors on the cells tested corresponded with the attachment of the viruses that were studied with respect to predicted receptor specificity. Virus attachment was not always predictive for efficient infection of the cells.
Rimmelzwaan
Guus F
GF
Department of Virology, Postgraduate School Molecular Medicine, Erasmus Medical Center, Rotterdam, The Netherlands. g.rimmelzwaan@erasmusmc.nl
Nieuwkoop
Nella J
NJ
de Mutsert
Gerrie
G
Boon
Adrianus C M
AC
Kuiken
Thijs
T
Fouchier
Ron A M
RA
Osterhaus
Albert D M E
AD
D000818:Animals; D001717:Birds / Q000821:virology; D002460:Cell Line; D005434:Flow Cytometry; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D009976:Orthomyxoviridae Infections / Q000821:virology*; D011991:Receptors, Virus / Q000378:metabolism; D053585:Virus Attachment; D014779:Virus Replication
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011991:Receptors, Virus