FIG 4.

Specificity of anti-FliD antibody and inhibition of Caco-2 cell binding by anti-FliD and anti-H40 antibodies. (A) The specific recognition of recombinant FliD protein by anti-FliD antibody was confirmed by immunoblotting. Note the intense reaction obtained with recombinant FliD protein and purified aEPEC 1711-4 native flagellin (PP), which is absent from the FliD-deficient mutant and drastically reduced in the FliC mutant. (B) Inhibition of adherence of strain 1711-4 to Caco-2 cells after incubation with anti-FliD or preimmune serum. (C) Inhibition of adherence of strain 1711-4 to Caco-2 cells by rabbit anti-FliC-H40 serum. Values were obtained from three independent experiments performed in triplicate. (D) The specificity of the anti-FliC-H40 serum was evaluated by immunostaining. Note the localization of colloidal gold nanoparticles on the flagellar structure (arrow). WT, wild type; SD, standard deviation.