Fluorescent poliovirus for flow cytometric cell surface binding studies

• Published in: Journal of virological methods Vol. 134; no. 1; pp. 1 - 7
• Main Authors: Freistadt, M.S., Eberle, K.E.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.06.2006; Amsterdam Elsevier; New York, NY
• Subjects: Biological and medical sciences; Cytometry; Flow Cytometry - methods; Fluorescence; Fluorescent Dyes - metabolism; Fundamental and applied biological sciences. Psychology; HeLa Cells - virology; Humans; Microbiology; Poliomyelitis - virology; Poliovirus; Poliovirus - metabolism; Poliovirus - physiology; Receptors; Staining and Labeling; Techniques used in virology; U937 Cells - virology; Virology; Virus Replication; Cytometry; Receptors; moi; Fluorescence; CNS; Poliovirus; pfu; MAb; Virus; Flow cytometry; Microbiology; Picornaviridae; Enterovirus; Method; Cell surface; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2005.08.011

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.