Construction and characterization of a replication-defective herpes simplex virus 2 ICP8 mutant strain and its use in immunization studies in a guinea pig model of genital disease.
A replication-defective mutant of herpes simplex virus 2 (HSV-2) was engineered by replacing the ICP8 gene of HSV-2 strain 186 with an ICP8-lacZ fusion gene from the herpes simplex virus 1 (HSV-1) HD-2 mutant strain. The resulting virus, HSV-2 5BlacZ, is defective for growth in Vero cells but is capable of growth in a cell line that expresses HSV-1 ICP8. In Vero cells, the mutant virus is defective for DNA synthesis but is able to express many viral proteins at levels similar to those of wild-type virus, including several of the late kinetic class. SDS-PAGE and Western blot analysis demonstrated the expression of glycoproteins B and D by 5BlacZ in Vero cells. Initial studies have shown that immunization with 5BlacZ protects guinea pigs from intravaginal HSV-2 challenge. Immunized animals had less severe genital skin disease and reduced replication of the challenge virus in the genital tract during primary infection and reduced episodes of recurrent disease. Thus, HSV-2 ICP8 shows gene regulatory properties similar to those of HSV-1 ICP8, and this HSV-2 ICP8 mutant virus shows a phenotype similar to those of HSV-1 ICP8 mutant strains. Replication-defective mutants of HSV-2 offer a potential vaccine approach for immune intervention against HSV-2 genital disease and latent infection.
Da Costa
X J
XJ
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Bourne
N
N
Stanberry
L R
LR
Knipe
D M
DM
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003673:Defective Viruses / Q000235:genetics*; D004195:Disease Models, Animal; D005260:Female; D016679:Genome, Viral; D006168:Guinea Pigs; D006558:Herpes Genitalis / Q000276:immunology / Q000517:prevention & control*; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000502:physiology; D007763:Lac Operon; D010641:Phenotype; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology / Q000627:therapeutic use; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology / Q000627:therapeutic use*
D014614:Vaccines, Synthetic; D014764:Viral Proteins; D014765:Viral Vaccines; C047419:herpes simplex virus type-2 protein ICP8