Significance of the secreted form of IpaC, a 45 kDa protein of Shigella dysenteriae 1, in the invasive process as determined by monoclonal antibodies.
Invasion plasmid antigen C (IpaC), a 45 kDa plasmid encoded protein, is associated with the virulence of virulent Shigella spp. In S. dysenteriae type 1 the 45 kDa IpaC protein is secreted to a greater extent into the surrounding medium in comparison to other Shigella spp. Monoclonal antibodies (mAbs) to the secreted form of IpaC protein were raised in this study. Of the four secretory hybrid cells, one (3G4) was found to have a very high antibody titre as determined by ELISA. The specificity of 3G4 was confirmed by immunoblotting of whole cell extract of Escherichia coli strain MC1061 carrying the plasmid pHW756 which synthesizes both the IpaB and C proteins. The effect of the mAbs on plaque formation by virulent Shigella dysenteriae 1 was determined and it was found that the clone 3G4 substantially (55%) reduced plaque formation on HeLa cell monolayer. The epitope specificity of the mAb 3G4 was competitively inhibited by the convalescent phase sera from human, suggesting that the epitope recognized by clone 3G4 was expressed during the natural course of infection and also indicating that the 45 kDa (IpaC) protein in secreted form has a definite role in the invasive process.
Shaikh
N M
NM
Division of Microbiol Genetics, National Institute of Cholera and Enteric Diseases, Calcutta, India.
Nair
G B
GB
Kumar
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000032:analysis / Q000096:biosynthesis* / Q000097:blood; D004405:Dysentery, Bacillary / Q000097:blood; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008970:Molecular Weight; D010957:Plasmids; D012762:Shigella dysenteriae / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D013045:Species Specificity; D014774:Virulence / Q000235:genetics
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; C092342:IpaC protein, Shigella