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J Virol. 1972 November; 10(5): 1021-1030
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

Isolation of a Herpes Simplex Virus-Specific Antigenic Fraction Which Stimulates the Production of Neutralizing Antibody

Gary H. Cohen, Manuel Ponce De Leon and Chris Nichols

1 Department of Microbiology, School of Dental Medicine, University of Pennsylvania, and the Center for Oral Health Research, Philadelphia, Pennsylvania 19104

ABSTRACT

Infection of mammalian cells with herpes simplex virus (HSV) results in the production of a number of virus-induced soluble antigens. Immunodiffusion analyses of the soluble antigen mixture (SAM) obtained from HSV-infected KB or BHK cells revealed at least six well-defined immunoprecipitin bands. Calcium phosphate chromatography (Brushite) was employed to separate one immunoprecipitin (designated CP-1) from the remaining viral and host antigens. We conclude that CP-1 is a viral-specific antigen because (i) specific antiserum, which had been repeatedly absorbed with uninfected cell extracts or serum components, still retained the capacity to react in gel diffusion with CP-1 antigen; (ii) anti-CP-1 serum reacted in gel diffusion with SAM, yielding one precipitin band in identity with the band formed against human gamma globulin; (iii) the CP-1 fraction stimulated the production of HSV-neutralizing antibody of high capacity. The last observation suggests that fraction CP-1 contains a biologically active structural component of the virus which is associated with the envelope. The CP-1 immunoprecipitin was separated from SAM by an alternative method by using a cyanogen bromide-linked immunosorbent prepared from anti-CP-1 gamma globulin. The observation that the CP-1 antigen isolated from the immunosorbent effectively blocked serum-neutralizing activity provided further evidence that neutralizing antibody was directed against CP-1. Acrylamide gel electrophoresis and immunological experiments suggest that the CP-1 antigen is in part a glycoprotein. The finding that CP-1 contains only one antigenic component of the virus will permit future biological studies to be made with a monoprecipitin antiserum. In addition, the techniques described in this paper represent initial steps in the purification of HSV antigens.


J Virol. 1972 November; 10(5): 1021-1030
Copyright © 1972 American Society for Microbiology. All Rights Reserved.




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Copyright © 1972 by the American Society for Microbiology. All rights reserved.