Inhibition of in vitro growth of coliform bacteria by a monoclonal antibody directed against ferric enterobactin receptor FepA.

The ability of a murine monoclonal antibody that blocks the enterobactin ligand-binding site of the ferric enterobactin receptor FepA to inhibit the growth of coliform bacteria derived from a bovine intramammary infection (IMI) was determined in an iron-restricted medium. Bacterial isolates from bovine IMI in five herds were tested by the chrome azurol sulfonate assay to detect siderophore production. Each of the isolates of Escherichia coli (n = 25) and Klebsiella pneumoniae (n = 25) were positive for siderophore production. Each isolate expressed iron-regulated outer membrane proteins when grown in trypticase soy broth plus the iron chelator alpha-alpha'-dipyridyl. Immunoblots revealed that the monoclonal antibody recognized FepA that was expressed by each of the E. coli isolates (n = 25). Only 4 of 25 K. pneumoniae isolates produced FepA that reacted with the monoclonal antibody. This result coincided with the results of an in vitro growth assay. Growth of all E. coli isolates was significantly inhibited by the addition of monoclonal antibody to synthetic medium containing apolactoferrin. Antigenic variation in the enterobactin-binding site resulted in a low percentage of K. pneumoniae isolates that were inhibited by the monoclonal antibody. Inhibition of bacterial growth by the monoclonal antibody was dose-dependent. As little as 50 micrograms/ml of purified antibody had an inhibitory effect on bacterial growth in the synthetic iron-restricted medium. Monoclonal antibody specific for the enterobactin ligand-binding site of FepA inhibited the growth of E. coli that was isolated from bovine IMI.