Antigenic analysis of Campylobacter flagellar protein and other proteins

Abstract

Outer membrane proteins of Campylobacterjejuni and other campylobacter species were analyzed for their antigenic potentials by immunoblotting. Polypeptides were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred electrophoretically, and reacted with rabbit antisera to C. jejuni. Each Campylobacter species analyzed demonstrated a unique outer membrane protein antigenic profile; interspecies antigen sharing was observed to be compatible with the degree of DNA relatedness between the species. The most highly conserved outer membrane protein antigen was the flagellum (molecular weight, 62,000). An aflagellate mutant was found to be untypable with the heat-labile system, in contrast to its parental isolate. The immunogenic potentials of C. jejuni proteins were examined by immunoblot analysis of sera from infected humans. Sera of convalescent patients, reacted with their homologous C. jejuni isolates, recognized a variety of campylobacter proteins. The most consistent immunogen in human infection was the flagellar protein. Patient sera assayed by the immunoblot technique were easily distinguished from control sera, which did not recognize specific campylobacter antigens. These findings suggest that the campylobacter flagellar protein is an essential determinant of the heat-labile antigen typing scheme and is the dominant immunogen recognized during C. jejuni infections in humans.

Occurrence of Campylobacter spp. in Young Gulls, Duration of Campylobacter Infection and Reinfection by Contact*

“…On the other hand, studies (2) suggest that there is an antigenetic similarity between the outer membrane proteins (OMP) of different C. jejuni strains and that some OMPs have vaccinogenic potential. The major OMP is an important intraspecies antigen (21), and an antigenetic similarity between flagellin proteins of thermophilic Campylobacters and a considerable antigenetic cross-reactivity of the major OMP has been demonstrated (13). Also cross-reactions of antisera and lipopolysaccharides between different Campyfobacter strains were noted in some cases (17).…”

Role of the 25 kDa major outer membrane protein ofLegionella pneumophilain attachment to U‐937 cells and its potential as a virulence factor for chick embryos

“…Outer membrane proteins for all Leg. pneumophila samples were isolated by methods previously described (Wenman et al 1985), assayed for protein content (Lowry et al 1951) and subjected to SDS-PAGE (Laemmli 1970). Gels were silverstained by the method described by Oakley et al (1980).…”

Purification and Antigenic Analysis of Flagella of Campylobacter jejuni

“…During the purification steps, two contaminating peptides, of 92,000 and 42,000 daltons, appeared Table 1. Slide agglutination of various strains of C. jejuni and C. coli by anti-flagellin antiserum against C. jejuni FUM 158432 as has been observed by others (3,28). Since the latter peptide could be removed by treating the flagellar preparation with Triton X-100 before applying it to the column of DEAE-Sephacel, it could be a peptide associated with a membranous structure, possibly with the outer membrane protein as stated by Logan and Trust (14).…”

“…The reports on the purification of the flagella of C. jejuni so far presented indicate Y. UEKI ET AL that some unknown peptides were usually associated with the flagellar preparation (3,14,28). Among these peptides the protein of 45,000 daltons was recently reported to be an outer membrane peptide (14).…”