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Immunoselection of recombinant baculoviruses expressing high levels of biologically active herpes simplex virus type 1 glycoprotein D

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Summary

The DNA sequence encoding the complete herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) was inserted into a baculovirus transfer vector under control of the polyhedrin gene promoter of the baculovirusAutographa california nuclear polyhedrosis virus (AcNPV). After co-transfection ofSpodoptera frugiperda (Sf9) insect cells with wild-type AcNPV DNA and the recombinant transfer vector DNA, polyhedrin-negative recombinants that expressed high levels of HSV-1 gD were isolated using immunoaffinity selection with antibody coated magnetic particles followed by plaque purification. These recombinant baculoviruses expressed a protein that was slightly smaller than virion HSV-1 gD made in Vero cells. This recombinant protein was expressed at high levels. The expressed protein was glycosylated, was found on the membrane of Sf9 cells, and reacted with gD specific antibodies. Antibodies raised in mice to the recombinant gD neutralized HSV-1 as measured by plaque reduction assays. Mice inoculated with the recombinant baculovirus were completely protected from lethal challenge with HSV-1.

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Authors and Affiliations

  1. Ophthalmology Research, Cedars-Sinai Medical Center, Los Angeles, California, U.S.A.

    Homayon Ghiasi, A. B. Nesburn, Ravi Kaiwar & S. L. Wechsler

  2. Department of Ophthalmology, University of California, Los Angeles, School of Medicine, Los Angeles, California, U.S.A.

    Homayon Ghiasi, A. B. Nesburn & S. L. Wechsler

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  1. Homayon Ghiasi
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  2. A. B. Nesburn
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  3. Ravi Kaiwar
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  4. S. L. Wechsler
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Ghiasi, H., Nesburn, A.B., Kaiwar, R. et al. Immunoselection of recombinant baculoviruses expressing high levels of biologically active herpes simplex virus type 1 glycoprotein D. Archives of Virology 121, 163–178 (1991). https://doi.org/10.1007/BF01316752

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  • DOI: https://doi.org/10.1007/BF01316752

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