Alpha-actinin accumulation in epithelial cells infected with attaching and effacing gastrointestinal pathogens.
Fluorescent F-actin staining utilizing phalloidin, a highly toxic mushroom poison, is used as an indirect test to detect attaching and effacing (AE) bacteria. A study was done to determine if accumulation of alpha-actinin in infected tissue culture cells is a consistent feature and whether it corresponds with the AE response. Rearrangement of alpha-actinin was detected using immunofluorescence microscopy by incubation of infected cells with a murine monoclonal anti-alpha-actinin antibody. Foci of alpha-actinin-specific fluorescence corresponding to areas of bacterial adhesion were detected by transmission electron microscopy in HEp-2 and gastric KATO-III cells infected with only those bacterial strains that formed AE lesions. Therefore, this study shows that alpha-actinin accumulation is a consistent, specific manifestation of the AE phenotype and forms the basis for the development of a safe alternative test for detecting AE bacteria.
Ismaili
A
A
Division of Gastroenterology, Hospital for Sick Children, Toronto, Ontario, Canada.
Philpott
D J
DJ
Dytoc
M T
MT
Soni
R
R
Ratnam
S
S
Sherman
P M
PM
D000185:Actinin / Q000032:analysis / Q000378:metabolism*; D000199:Actins / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D001422:Bacterial Adhesion; D002460:Cell Line; D004848:Epithelium / Q000382:microbiology / Q000648:ultrastructure; D004926:Escherichia coli / Q000472:pathogenicity / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D005767:Gastrointestinal Diseases / Q000382:microbiology; D016998:Helicobacter / Q000472:pathogenicity / Q000502:physiology*; D016480:Helicobacter pylori / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D051379:Mice / Q000276:immunology; D008854:Microscopy, Electron; D010590:Phalloidine; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000199:Actins; D000911:Antibodies, Monoclonal; D000185:Actinin; D010590:Phalloidine