Selection of receptor-binding variants of human influenza A and B viruses in baby hamster kidney cells
| Autor: | N. V. Bovin, Mikhail Matrosovich, Elena A. Govorkova, C. Gerdil, B. Fanget, Alexander B. Tuzikov, Robert G. Webster |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Models
Molecular Human influenza viruses Sialoglycoproteins Mutant Hemagglutinin (influenza) Hemagglutinin Glycoproteins Influenza Virus Chick Embryo Kidney Dogs Virology Cricetinae Baby hamster kidney cell Animals Humans Protein Isoforms Receptor Cells Cultured Ovum chemistry.chemical_classification biology urogenital system Embryonated Embryo Flow Cytometry Amino acid Influenza B virus chemistry Amino Acid Substitution Influenza A virus Mutation biology.protein Receptors Virus |
| Zdroj: | Virology. 262(1) |
| ISSN: | 0042-6822 |
| Popis: | Cultivation of human influenza viruses in the allantoic cavity of embryonated chicken eggs leads to a selection of receptor-binding variants with amino acid substitutions on the globular head of the hemagglutinin (HA) molecule. Such selection can be avoided by growing the human viruses in Madin Darby canine kidney (MDCK) cells. In the present study, we tested whether baby hamster kidney (BHK) cells select receptor-binding mutants of human influenza viruses. After isolating H1N1, H3N2, and type B influenza viruses from clinical samples in MDCK cells, we passaged them in either BHK cells or chicken eggs. The BHK-grown viruses differed from their MDCK-grown counterparts by virtue of mutations in the HA: 225D → G (H1N1 virus), 128T → A and 226I → V (H3N2), and 187N → D (type B) (H3 numbering). Variants with different substitutions were selected by passaging of the same MDCK-grown parents in eggs: 141L → H, 208R → H, and 225D → G (H1N1), 194L → I (H3N2), and 137G → R (B). Compared with their MDCK-grown counterparts, both BHK- and egg-grown viruses possessed a higher affinity for the cellular membranes of BHK cells and of the chorioallantoic cells of chicken embryos and for a 3′-sialylgalactose-containing synthetic sialylglycopolymer. By contrast, changes in the affinity of mutants for a 6′-sialyl-( N -acetyllactosamine)-containing sialylglycopolymer varied from negative to positive. Fluorescence-activated cell-sorting analysis with linkage-specific lectins showed that the density of the 6′-sialyl-( N -acetyllactosamine)-containing receptors is substantially lower on the surface of BHK cells than on MDCK cells, providing an explanation for the growth restriction of human viruses in the former cells. Our data demonstrate that cultures of BHK cells, like eggs, can select receptor-binding variants of human influenza viruses. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect