Abstract
The antibody response of 30 volunteers vaccinated with a complex of group B polysaccharide and outer membrane vesicles (OMV) from serotype 2a Neisseria meningitidis and of 3 individuals who received a placebo vaccine was determined by immunoblotting. OMV were separated by sodium dodecyl sulfate-gel electrophoresis and electrotransferred to nitrocellulose filters. Binding of immunoglobulin G (IgG), IgA, and IgM antibodies in the human sera to OMV components was detected with class-specific peroxidase-conjugated antibodies. The immunoblotting results were also related to the bactericidal activity of the sera and the meningococcal carrier status of the volunteers. Before vaccination weakly reactive bands in the molecular weight range of 140,000 to 10,000 were observed on the blots. Sera from carriers showed more marked bands. Individual patterns of increased reactivity were seen 6 weeks after vaccination. The main immunoreactive components of OMV corresponded to a molecular weight of 43,000 (class 1 protein), 30,000 (class 5 proteins), and 22,000. IgG antibodies in postvaccination sera of high bactericidal titers showed distinct binding to the 43,000-molecular-weight antigen. Meningococcal carriers had antibodies against an antigen of 22,000 molecular weight; in polyacrylamide gels this component did not stain with Coomassie brilliant blue or silver. The marked binding of IgG antibodies to the class 5 proteins decreased considerably between weeks 6 and 25 after vaccination. Periodate oxidation of OMV abolished the binding of IgG antibodies to the class 5 proteins, whereas the antigenicity of the 43,000-molecular-weight (class 1 protein) and 22,000-molecular-weight antigens was unaffected.
MeSH Terms
Antibodies, Bacterial/analysis
Antibody Formation
Bacterial Outer Membrane Proteins/analysis
Bacterial Vaccines/immunology
Enzyme-Linked Immunosorbent Assay
Humans
Immunoglobulin A/analysis
Immunoglobulin G/analysis
Immunoglobulin M/analysis
Meningococcal Vaccines
Molecular Weight
Oxidation-Reduction
Periodic Acid/pharmacology
Time Factors
Vaccination
Chemicals
Antibodies, Bacterial
Bacterial Outer Membrane Proteins
Bacterial Vaccines
Immunoglobulin A
Immunoglobulin G
Immunoglobulin M
Meningococcal Vaccines
Periodic Acid
metaperiodate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wedege E
Frøholm L O
References (22)
22 references, click to expand
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
An outer membrane protein of Neisseria meningitidis group B responsible for serotype specificity.
J Exp Med. 1974 Jul 1;140(1):87-104
PMID: 4134571
-
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
PMID: 388439
-
Five structural classes of major outer membrane proteins in Neisseria meningitidis.
J Bacteriol. 1981 Apr;146(1):69-78
PMID: 6783622
-
Variability of low-molecular-weight, heat-modifiable outer membrane proteins of Neisseria meningitidis.
Infect Immun. 1980 Dec;30(3):642-8
PMID: 6785230
-
Alloantibodies to factor IX in Haemophilia B characterized by crossed immunoelectrophoresis and enzyme-conjugated antisera to human immunoglobulins.
Br J Haematol. 1981 May;48(1):15-23
PMID: 6788066
-
A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels.
Anal Biochem. 1982 Jan 1;119(1):115-9
PMID: 6176137
-
Protection against group B Neisseria meningitidis disease: preparation of soluble protein and protein-polysaccharide immunogens.
Infect Immun. 1982 Jul;37(1):271-80
PMID: 6809629
-
Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to a positively charged membrane filter.
Anal Biochem. 1982 Aug;124(2):396-405
PMID: 7149237
-
Immunogenicity of meningococcal antigens as detected in patient sera.
Infect Immun. 1983 Apr;40(1):398-406
PMID: 6131872
-
Heterogeneity and variation among Neisseria meningitidis lipopolysaccharides.
J Bacteriol. 1983 Aug;155(2):498-504
PMID: 6409879
-
Monoclonal antibody that recognizes an outer membrane antigen common to the pathogenic Neisseria species but not to most nonpathogenic Neisseria species.
Infect Immun. 1984 Mar;43(3):994-9
PMID: 6421742
-
Use of a zwitterionic detergent for the restoration of the antibody-binding capacity of electroblotted meningococcal outer membrane proteins.
J Immunol Methods. 1984 Feb 24;67(1):1-11
PMID: 6421934
-
Meningococcal group B vaccine trial in Norway 1981--1982. Preliminary report of results available November 1982.
NIPH Ann. 1983 Dec;6(2):133-8
PMID: 6427703
-
An ELISA study of the antibody response after vaccination with a combined meningococcal group B polysaccharide and serotype 2 outer membrane protein vaccine.
NIPH Ann. 1983 Dec;6(2):139-49
PMID: 6427704
-
Immunoblotting and dot immunobinding--current status and outlook.
J Immunol Methods. 1984 Sep 4;72(2):313-40
PMID: 6206159
-
Analyses of gonococcal lipopolysaccharide in whole-cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: stable association of lipopolysaccharide with the major outer membrane protein (protein I) of Neisseria gonorrhoeae.
Infect Immun. 1984 Oct;46(1):202-12
PMID: 6207109
-
Monoclonal antibodies to serotype 2 and serotype 15 outer membrane proteins of Neisseria meningitidis and their use in serotyping.
Infect Immun. 1984 Oct;46(1):260-6
PMID: 6434428
-
Naturally occurring antibodies in human sera that react with the iron-regulated outer membrane proteins of Escherichia coli.
Infect Immun. 1985 Mar;47(3):808-13
PMID: 2982739
-
Recognition of serogroup A Neisseria meningitidis serotype antigens by human antisera.
Infect Immun. 1985 Apr;48(1):23-8
PMID: 3920147
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
THE PERIODATE OXIDATION OF AMINO ACIDS WITH REFERENCE TO STUDIES ON GLYCOPROTEINS.
Biochem J. 1965 Jan;94:17-24
PMID: 14342227