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The Detection of Epstein—Barr Virus Receptors Utilizing Radiolabelled Virus

Norio Koide

Department of Tumor Biology, Karolinska Institutet, S-104 01 Stockholm, Sweden

Epstein—Barr virus (EBV) was labelled with ³H-thymidine and purified about 1000-fold from the culture medium by ultracentrifugation on 5 to 30% dextran grandients. The presence of the virus was monitored by radioactivity and Epstein—Barr virus-determined nuclear antigen (EBNA) induction in sensitive indicator cells (Ramos). Peaks for both activities occurred in the 17 to 18% dextran fractions. Unlabelled virus recovered in the peak fraction was labelled with ¹²⁵I. Both thymidine and ¹²⁵I-labelled purified virus bound quantitatively to receptor-positive Burkitt lymphoma-derived cell lines but not to EBV-receptor-negative T-lymphocyte-derived cell lines. Thymidine-labelled virus that was allowed to bind to Raji cells was present in the interior of briefly trypsinized cells after 3 h incubation at 37 °C. The results provide a convenient method for detecting the EBV receptor by radioactively labelled virus.

Present address: Department of Internal Medicine, Okayama University School of Medicine, 5-1, 2-chome, Okayama 700, Japan.

Received 29 August 1980; accepted 7 January 1981.