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Application of Immunohistochemistry to Study of Avian Leukosis Virus

Departments of Microbiology and Anatomy, State University of New York, Upstate Medical Center, Syracuse, New York, 13210, U.S.A.

A modification of the peroxidase-labelled antibody technique was applied to study the distribution of virus antigens in chicken cells infected with two serotypes (A and B) of avian leukosis virus (ALV). Type-specific chicken antisera reacted only with cells infected with virus of the homologous envelope serotype. When unfixed cells were exposed to type-specific antivirus serum, only antigens located at the cell surface were stained, while cells exposed to type-specific antibodies after fixation revealed both surface and intracytoplasmic virus antigen. Cytoplasmic antigen was usually concentrated in discrete granules which often had a vesicular structure.

Hamster antibodies against ALV group-specific (gs) antigen reacted with fixed infected cells regardless of envelope serotype, and the distribution of antigen was as shown with chicken type-specific antibodies. Intranuclear gs antigen may have been present in a few cells. Unfixed infected cells did not react with hamster gs antibody and confirmed the location of ALV gs antigen within the virus particle.

Some chicken antisera with neutralizing antibodies against a single envelope serotype of ALV contained both type-specific and gs antibodies. These antisera reacted with fixed cells infected with virus of either the homologous or heterologous serotype and stained both surface and cytoplasmic antigens. With unfixed infected cells, these antisera combined only with the surface of cells infected with virus of the same serotype as the neutralizing antibodies in the serum. Thus, the reaction in heterologously infected fixed cells was with internal gs antigen. This confirms independently that chickens are not naturally tolerant to their homologous ‘C type’ gs antigens.

Received 6 September 1971; accepted 3 January 1972.