Fluorescent poliovirus for flow cytometric cell surface binding studies.
Specific cell-surface binding is the essential first step for cellular invasion by viruses. To understand this process, various methods to evaluate binding properties of viruses to cells have been developed. However, many rely on radioactive labeling or indirect immunofluorescence. The development of a novel fluorescence binding assay for poliovirus is described. Poliovirus (type 1 Mahoney or Sabin) was labeled directly with fluorescein using a commercially available fluoresceination kit. Fluorescently labeled poliovirus was bound to its specific receptor on Hela or U937 cells and detected by flow cytometric analysis. Specific binding and infectivity was retained, although reduced, depending on the extent of fluoresceination. Therefore, depending on the users' requirements, the extent of fluoresceination must be titrated carefully to achieve maximal fluorescence and minimal functional destruction. It is likely that this method may be useful with other viruses.
Freistadt
M S
MS
Department of Microbiology, Immunology and Parasitology, LSU Health Sciences Center, Box P6-1, 1901 Perdido St., New Orleans, LA 70112, USA. mfreis@lsuhsc.edu
Eberle
K E
KE
D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D011051:Poliomyelitis / Q000821:virology; D017955:Poliovirus / Q000378:metabolism / Q000502:physiology*; D013194:Staining and Labeling; D020298:U937 Cells / Q000821:virology; D014779:Virus Replication
D005456:Fluorescent Dyes