Vaccinia virus entry/fusion complex subunit A28 is a target of neutralizing and protective antibodies.
The vaccinia virus entry/fusion complex (EFC) is comprised of at least eight transmembrane proteins that are conserved in all poxviruses. However, neither the physical structure of the EFC nor the immunogenicity of the individual components has been determined. We prepared soluble forms of two EFC components, A28 and H2, by replacing the transmembrane domain with a signal peptide and adding a polyhistidine tail. The proteins were expressed by baculoviruses, secreted from insect cells, purified by affinity chromatography and used to raise antibodies in rabbits. The antibodies recognized the viral proteins but only the antibody to recombinant A28 bound intact virions and neutralized infectivity. Analyses with a set of overlapping peptides revealed a neutralizing epitope between residues 73 and 92 of A28. Passive immunization of mice with IgG purified from the anti-A28 serum provided partial protection against a vaccinia virus intranasal challenge, whereas IgG from the anti-H2 serum did not.
Nelson
Gretchen E
GE
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-3210, USA.
Sisler
Jerry R
JR
Chandran
Dev
D
Moss
Bernard
B
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology* / Q000627:therapeutic use; D016367:Baculoviridae / Q000235:genetics; D001835:Body Weight; D002846:Chromatography, Affinity; D018604:Epitope Mapping; D005260:Female; D015870:Gene Expression; D007074:Immunoglobulin G / Q000276:immunology / Q000627:therapeutic use; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014615:Vaccinia / Q000517:prevention & control; D014616:Vaccinia virus / Q000276:immunology* / Q000502:physiology*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*; D053586:Virus Internalization
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins
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