Immunobiology of purified recombinant outer membrane porin protein I of Neisseria gonorrhoeae.
Gonococcal porins (Por) from strains FA19 (Por-1, serogroup A), MS11 (Por-2, serogroup B) and FA6434 (Por-5, a hybrid porin containing epitopes from both serogroups), were expressed in Escherichia coli and purified under non-denaturing conditions. Porins were inserted into liposomes, and they were bound by monoclonal antibodies which bind native Por and intact gonococci, but not denatured Por. All three recombinant porins (rPor) were highly immunogenic in rabbits without additional adjuvant. The rPor antisera were specific for Por by Western blotting and whole-cell radioimmunoprecipitation and were broadly cross-reactive within serogroups. Post-immune, but not pre-immune, sera bound to intact gonococci, induced deposition of complement components C3 and C9 onto gonococcal membranes and increased association with and activation of human neutrophils. Gonococci were not killed in bactericidal assays, and there was no phagocytic killing with gonococci opsonized with recombinant antisera. Lack of killing in bactericidal assays was not caused by the presence of blocking antibodies to the outer-membrane protein Rmp.
Elkins
C
C
Department of Medicine, School of Medicine, University of North Carolina, Chapel Hill 27599.
Barkley
K B
KB
Carbonetti
N H
NH
Coimbre
A J
AJ
Sparling
P F
PF
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000145:classification / Q000235:genetics / Q000276:immunology*; D018272:Porins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012703:Serotyping
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D018272:Porins; D011994:Recombinant Proteins; C057699:porin protein, Neisseria