Recombinant anti-EspA antibodies block Escherichia coli O157:H7-induced attaching and effacing lesions in vitro

• DOI: 10.1016/j.micinf.2005.07.009
• Publication Date: 2005-09-20T13:03:55Z
• Authors: Roberto M. La Rag...; Shila Patel; Ben Maddison; Martin J. Woodward; Angus Best; Garry C. Whitelam; Kevin C. Gough
• Keywords: 0303 health sciences; Staining and Labeling; Phalloidine; Escherichia coli Proteins; Escherichia coli O157; Antibodies, Bacterial; Actins; Bacterial Adhesion; Recombinant Proteins; Cell Line; 3. Good health; 03 medical and health sciences; Animals; Humans; Rabbits; Immunoglobulin Fragments; Epitope Mapping; Fluorescein-5-isothiocyanate

Intimin and EspA proteins are virulence factors expressed by attaching and effacing Escherichia coli (AEEC) such as enteropathogenic and enterohaemorrhagic E. coli. The EspA protein makes up a filament structure forming part of the type III secretion system (TTSS) that delivers effector proteins to the host epithelial cell. Bacterial surface displayed intimin interacts with translocated intimin receptor in the host cell membrane leading to intimate attachment of the bacterium and subsequent attaching and effacing lesions. Here, we have assessed the use of recombinant monoclonal antibodies against E. coli O157:H7 EspA and intimin for the disruption of AEEC interaction with the host cell. Anti-gamma intimin antibodies did not reduce either adhesion of E. coli O157:H7 to host cell mono-layers or subsequent host cell actin rearrangement. Anti-EspA antibodies similarly had no effect on bacterial adhesion however they had a marked effect upon E. coli O157:H7-induced host cell actin rearrangement, where both monoclonal and polyclonal antibodies completely blocked cytoskeletal changes within the host cell. Furthermore, these anti-EspA antibodies were shown to reduce actin rearrangement induced by some but not all other AEEC serotypes tested. Both polyclonal and monoclonal antibodies could be used to label E. coli O157 EspA filaments and these immunoreagents did not inhibit the formation of such filaments. This is the first report of monoclonal antibodies to EspA capable of disrupting the TTSS function of E. coli O157:H7.