Measurement of bluetongue virus binding to a mammalian cell surface receptor by an in situ immune fluorescent staining technique.
A quantifiable in situ immune fluorescent assay (IFA) was developed to measure bluetongue virus (BTV) binding to mammalian cells. The utility of the assay was demonstrated with both Chinese hamster ovary (CHO) and bovine pulmonary artery endothelial (CPAE) cells. Since heparin sulfate (HS) has been shown to function as a receptor for a number of viruses, its role as a receptor for BTV was evaluated with the in situ IFA. Binding of BTV to both CHO and CPAE cells was inhibited in a dose dependent manner by HS. In addition, HS deficient CHO cells showed greatly diminished binding of BTV when compared to the parental cell line. The IFA protocol will find application, as a non-isotopic, quantifiable technique, to study virus-cell receptor interactions. Information gained from such studies will expand our understanding of the early steps in virus replication.
Mecham
James O
JO
USDA, ARS, Arthropod-Borne Animal Diseases Research Laboratory, 1000 E University Ave., Laramie, WY 82071, USA. james.mecham@ars.usda.gov
McHolland
Linda E
LE
D000818:Animals; D001820:Bluetongue virus / Q000502:physiology*; D002417:Cattle; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods; D006497:Heparan Sulfate / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000378:metabolism*; D011991:Receptors, Virus / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000378:metabolism*; D014773:Virology / Q000379:methods*; D053585:Virus Attachment
D006497:Heparan Sulfate; D011991:Receptors, Virus