Studies of the Epstein-Barr virus receptor found on Raji cells. I. Extraction of receptor and preparation of anti-receptor antibody
Simmons, J.G.; Hutt-Fletcher, L.M.; Fowler, E.; Feighny, R.J.
Journal of Immunology 130(3): 1303-1308
1983
ISSN/ISBN: 0022-1767 PMID: 6337215 Document Number: 218906
Raji, a human lymphoblastoid cell line, expresses a membrane receptor (EBVR) specific for Epstein Barr virus (EBV). A component that binds EBV was extracted from this cell line by treatment of the cells for 3 h on ice with Tris buffer containing 10% glycerol. The treatment reduce the capacity of the cells to bind virus, and after concentration the receptor extract (RE) inhibited both EBV binding and superinfection of fresh Raji cells. Similar prepared extracts of EBVR- cells lacked such activity. An antibody was made to the extract (anti-RE), which, after absorption with EBVR- cells, bound to the same percentages of EBVR+ lymphoblastoid cell lines, EBVR+ human/mouse somatic cell hybrids, and fresh peripheral B cells as the virus did. In reciprocal assays, preincubation of EBVR+ cells with anti-RE inhibited virus binding. Doubly stained patches were observed on membranes of EBVR+ cells that had been incubated simultaneously with virus and anti-RE and stained respectively with rhodaminated and fluoresceinated reagents. The major polypeptide immunoprecipitated by anti-RE from radiolabeled Raji cells had no approximate calculated MW of 150,000.