title: Protective immunity to vaccinia virus induced by vaccination with multiple recombinant outer membrane proteins of intracellular and extracellular virions.
abstract: Infectious intracellular and extracellular forms of vaccinia virus have different outer membrane proteins, presenting multiple targets to the immune system. We investigated the immunogenicity of soluble forms of L1, an outer membrane protein of the intracellular mature virus, and of A33 and B5, outer membrane proteins of the extracellular enveloped virus. The recombinant proteins, in 10-microg amounts mixed with a Ribi- or saponin-type adjuvant, were administered subcutaneously to mice. Antibody titers to each protein rose sharply after the first and second boosts, reaching levels that surpassed those induced by percutaneous immunization with live vaccinia virus. Immunoglobulin G1 (IgG1) antibody predominated after the protein immunizations, indicative of a T-helper cell type 2 response, whereas live vaccinia virus induced mainly IgG2a, indicative of a T-helper cell type 1 response. Mice immunized with any one of the recombinant proteins survived an intranasal challenge with 5 times the 50% lethal dose of the pathogenic WR strain of vaccinia virus. Measurements of weight loss indicated that the A33 immunization most effectively prevented disease. The superiority of protein combinations was demonstrated when the challenge virus dose was increased 20-fold. The best protection was obtained with a vaccine made by combining recombinant proteins of the outer membranes of intracellular and extracellular virus. Indeed, mice immunized with A33 plus B5 plus L1 or with A33 plus L1 were better protected than mice immunized with live vaccinia virus. Three immunizations with the three-protein combination were necessary and sufficient for complete protection. These studies suggest the feasibility of a multiprotein smallpox vaccine.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Christiana
authors.initials: C
authors.lastname: Fogg
authors.forename: Shlomo
authors.initials: S
authors.lastname: Lustig
authors.forename: J Charles
authors.initials: JC
authors.lastname: Whitbeck
authors.forename: Roselyn J
authors.initials: RJ
authors.lastname: Eisenberg
authors.forename: Gary H
authors.initials: GH
authors.lastname: Cohen
authors.forename: Bernard
authors.initials: B
authors.lastname: Moss
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D017358:Cell Wall Skeleton / Q000008:administration & dosage; D003311:Cord Factors / Q000008:administration & dosage; D005260:Female; D007074:Immunoglobulin G / Q000097:blood; D008050:Lipid A / Q000008:administration & dosage / Q000031:analogs & derivatives*; D008562:Membrane Glycoproteins / Q000008:administration & dosage / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology; D014611:Vaccination; D017778:Vaccines, Combined / Q000276:immunology; D022223:Vaccines, Subunit / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014615:Vaccinia / Q000517:prevention & control*; D014616:Vaccinia virus / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000008:administration & dosage / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000008:administration & dosage / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*; D015431:Weight Loss
chemical_list: C446940:A33R protein, Vaccinia virus; D000914:Antibodies, Viral; D017358:Cell Wall Skeleton; D003311:Cord Factors; D007074:Immunoglobulin G; C087715:L1R protein, Vaccinia virus; D008050:Lipid A; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C075897:Ribi adjuvant; D017778:Vaccines, Combined; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D014765:Viral Vaccines; C074512:42kDa protein, Vaccinia virus