Variation within serovars of Neisseria gonorrhoeae detected by structural analysis of outer-membrane protein PIB and by pulsed-field gel electrophoresis
Outer-membrane protein PI is the antigen responsible for serovar specificity of Neisseria gonorrhoeae and is a potential vaccine target. In order to investigate possible hidden variation within a serovar, the sequence of the por genes encoding protein PIB have been obtained from a series of strains, including isolates known to be epidemiologically linked. The inferred amino acid sequences of the PIB molecules of isolates from known sexual contacts were identical, but non-related isolates showed significant heterogeneity in PIB sequence. These differences were not confined to the two variable regions (Var1 and Var2) which have previously been identified, but were largely, although not exclusively, located in regions predicted to form one of eight surface-exposed loops. The isolates were subjected to pulsed-field gel electrophoresis of restriction digests of chromosomal DNA, which also demonstrated identity between linked strains but revealed diversity within a serovar. The deduced amino acid sequences of PIB were also used to synthesize peptides for epitope-mapping experiments. These revealed that some mAbs, used to define serovar specificity, recognized linear epitopes located in loops 5 and 6, while others appeared to recognize conformational epitopes elsewhere in the molecule. The occurrence of the sequence differences within a serovar, which are not detected by the serotyping reagents, reveals that PIB represents a potential source of information which should permit considerably more detailed epidemiological studies than are currently possible and focuses attention on more conserved regions of the protein as potential targets for vaccination.
AndersonR. M.,
GuptaS.,
NgW.1990; The significance of sexual partner contact networks for the transmission dynamics of HIV.. J Acquired lmmune Defic Syndr 2:417–419
BrooksJ. L.,
FallonR. J.,
HeckelsJ. E.1995; Sequence variation in class 1 outer membrane protein in Neisseria meningitidis isolated from patients with meningococcal infection and close household contacts.. FEMS Microbiol Lett 128:145–150
ButtN. J.,
LambdenP. R.,
HeckelsJ. E.1990a; The nucleotide sequence of por gene encoding expression of outer membrane protein PIB in Neisseria gonorrhoeae strain P9.. Nucleic Acids Res 14:4258
ButtN. J.,
VirjiM.,
VayredaF.,
LambdenP. R.,
HeckelsJ. E.1990b; Gonococcal outer-membrane protein PIB: comparative sequence analysis and localization of epitopes which are recognized by type-specific and cross-reacting monoclonal antibodies.. J Gen Microbiol 136:2165–2172
CarbonettiN. H.,
SparlingP. F.1987; Molecular cloning and characterization of the structural gene for protein I, the major outer membrane protein of Neisseria gonorrhoeae.. Proc Natl Acad Sci USA 84:9084–9088
CarbonettiN. H.,
SimnadV. I.,
SeifertH. S.,
SoM.,
SparlingP. F.1988; Genetics of protein I of Neisseria gonorrhoeae: construction of hybrid porins.. Proc Natl Acad Sci USA 85:6841–6845
ElkinsC.,
CarbonettiN. H.,
VarelaV. A.,
StirewaltD.,
KlapperD. G.,
SparlingP. F.1992; Antibodies to N-terminal peptides of gonococcal porin are bactericidal when gonococcal lipopolysaccharide is not sialylated.. Mol Microbiol 6:2617–2628
GotschlichE. C.,
SeiffM. E.,
BlakeM. S.,
KoomeyM.1987; Porin protein of Neisseria gonorrhoeae: cloning and gene structure.. Proc Nut1 Acad Sci USA 84:8135–8139
HeckelsJ. E.,
VirjiM.,
TinsleyC. R.1990; Vaccination against gonorrhoeae: the potential protective effect of immunization with a synthetic peptide containing a conserved epitope of gonococcal outer membrane protein IB.. Vaccine 8:225–230
IsonC. A.,
WhitakerL.,
RentonA.1992; Concordance of auxotype/serovar classes of Neisseria gonorrhoeae between sexual contacts.. Epidemiol Infect 109:265–271
KnappJ. S.,
TamM. R.,
NowinskiR. C.,
HolmesK. K.,
SandstromE. G.1984; Serological classification of Neisseria gonorrhoeae with use of monoclonal antibodies to gonococcal outer membrane protein I.. J lnfect Dis 50:44–48
KnappJ. S.,
HolmesK. K.,
BoninP.,
HookE. W.1987; Epidemiology of gonorrhoea: distribution and temporal changes in auxotype/serovar classes of Neisseria gonorrhoeae.. Sex Transm Dis 14:26–32
McGuinnessB. T.,
LambdenP. R.,
HeckelsJ. E.1993; Class 1 outer membrane protein of Neisseria rneningitidis: epitope analysis of the antigenic diversity between strains, implications for subtype definition and molecular epidemiology.. Mol Microbiol 7:505–514
MaidenM. C. J.,
BygravesJ. A.,
McCarvilJ.,
FeaversI. M.1992; Identification of meningococcal serosubtypes by polymerase chain reaction.. J Clin Microbiol 30:2835–2841
MeeB. J.,
ThomasH.,
CookeS. J.,
LambdenP. R.,
HeckelsJ. E.1993; Structural comparison and epitope analysis of outermembrane protein PIA from strains of Neisseria gonorrhoeae with differing serovar specificities.. J Gen Microbiol 139:2613–2620
O’RourkeM.,
IsonC. A.,
RentonA. M.,
SprattB. G.1995; Opa-typing: a high resolution tool for studying the epidemiology of gonorrhoea.. Mol Microbiol 17:865–875
SandstromE. G.,
ChenK. C. S.,
BuchananT. M.1982; Serology of Neisseria gonorrhoeae: co-agglutination serogroups WI and WII/III correspond to different outer membrane protein molecules.. Infect Immun 38:462–470
SukerJ.,
FeaversI. M.,
AchtmanM.,
MorelliG.,
WangJ. F.,
MaidenM. C. J.1994; The porA gene in serogroup A meningococci: evolutionary stability and mechanism of genetic variation.. Mol Microbiol 12:253–265
Van der LeyP.,
HeckelsJ. E.,
VirjiM.,
HoogerhoutP.,
PoolmanJ. T.1991; Topology of outer membrane porins in pathogenic Neisseria spp.. Infect Immun 59:2963–2971
VirjiM.,
ZakK.,
HeckelsJ. E.1986; Monoclonal antibodies to gonococcal outer membrane protein IB: use in investigation of the potential protective effect of antibodies directed against conserved and type-specific epitopes.. J Gen Microbiol 132:1621–1629
VirjiM.,
FletcherJ. N.,
ZakK.,
HeckelsJ. E.1987; The potential protective effect of monoclonal antibodies to gonococcal outer membrane protein IA.. J Gen Microbiol 133:2639–2646
ZakK.,
DiazJ. L.,
JacksonD.,
HeckelsJ. E.1984; Antigenic variation during infection with Neisseria gonorrhoeae: detection of antibodies to surface proteins in sera of patients with gonorrhea.. J Infect Dis 149:166–173
Variation within serovars of Neisseria gonorrhoeae detected by structural analysis of outer-membrane protein PIB and by pulsed-field gel electrophoresis