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Cross-Neutralization of Infectious Mononucleosis and Burkitt Lymphoma Strains of Epstein-Barr Virus with Hyperimmune Rabbit Antisera¹

From the Howard Hughes Medical Institute Laboratory in Departments of Pediatrics and Epidemiology and Public Health, Yale University School of Medicine, Room 405 LCI, 333 Cedar Street, New Haven, Connecticut 06510.

Abstract

Antisera that neutralize Epstein-Barr virus (EBV) were prepared in rabbits. Neutralization tests were based on inhibition of transformation of normal lymphocytes into cell lines or on inhibition of induction of early antigen. Supernatant culture fluids of human and marmoset lymphoblastoid cell lines productive of virus were the source of antigen. Neutralizing activity was not induced by immunization of rabbits with medium from EBV transformed cell lines that did not release virus. However, neutralizing activity appeared in the sera of rabbits inoculated with supernatant fluid of a producer line from which nearly all the virions had been removed by ultracentrifugation. Rabbit antisera with neutralizing activity reacted with radioiodinated virus purified by velocity sedimentation on a sucrose gradient.

Neutralizing capacity of the sera was not removed by absorption with primary human or marmoset leukocytes, by absorption with EBV transformed human B lymphoid cells that were not virus producers, nor by absorption with marmoset lymphoblastoid T cell lines transformed by Herpes saimiri virus. However, absorption of the antisera with EB virus producer lines caused a marked reduction or elimination of neutralizing antibody titer.

Cross-neutralization and cross-absorption tests did not indicate the existence of distinct serotypes. Antiserum prepared against the Hawley EBV strain from a patient with mononucleosis neutralized two viruses of Burkitt lymphoma origin, HR-1 and Olare. Antiserum raised against the nontransforming HR-1 strain neutralized the two transforming strains. Absorption of the sera with virus producer cells removed neutralizing activity directed against both homologous and heterologous viruses. The results indicate that EB viruses of diverse origin and diverse biologic properties share antigens, presumably located both on the viral envelope and on the cytoplasmic membrane of producer cell lines, which elicit neutralizing activity.

Footnotes

¹ This work was supported by grants from the United States Public Health Service CA 12055, CA 16038, AI 11611, and from the American Cancer Society VC-107.

² Address for further correspondence.