Antigen detection in enteropathogenic Escherichia coli using secretory immunoglobulin A antibodies isolated from human breast milk.
Enteropathogenic Escherichia coli (EPEC) produces a characteristic attaching and effacing (A/E) lesion in the small intestines of infected children. The immune response to EPEC infection remains poorly characterized. The molecular targets that elicit protective immunity against EPEC disease are unknown. In this study protein antigens from EPEC were identified using secretory immunoglobulin A (sIgA) antibodies isolated from milk from Mexican women by Western blot analysis. Purified sIgA antibodies, which inhibit the adherence of EPEC to cells, reacted to many EPEC proteins, the most prominent of which were intimin (a 94-kDa outer membrane protein) and two unknown proteins with apparent molecular masses of 80 and 70 kDa. A culture supernatant protein of 110 kDa also reacted strongly with the sIgA antibodies. The molecular size of this protein and its reactivity with specific anti-EspC antiserum suggest that it is EPEC-secreted protein C (EspC). These EPEC surface protein antigens were consistently recognized by all the different sIgA samples obtained from 15 women. Screening of clinical isolates of various O serogroups from cases of severe infantile diarrhea revealed that all EPEC strains able to produce the A/E lesion showed expression of intimin and the 80- and 70-kDa proteins. Such proteins reacted strongly with the purified sIgA pool. Moreover, nonvirulent E. coli strains were unable to generate a sIgA response. The immunogenic capacities of the 80- and 70-kDa proteins as virulence antigens have not been previously reported. The strong sIgA response to intimin and the 80- and 70-kDa proteins obtained in this study indicates that such antigens stimulate intestinal immune responses and may elicit protective immunity against EPEC disease.
Manjarrez-Hernandez
H A
HA
Departamento de Salud Publica, Facultad de Medicina, Universidad Nacional Autonoma de Mexico, Mexico D.F. 04510, Mexico.
Gavilanes-Parra
S
S
Chavez-Berrocal
E
E
Navarro-Ocaña
A
A
Cravioto
A
A
D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000032:analysis; D002478:Cells, Cultured; D004926:Escherichia coli / Q000276:immunology*; D005260:Female; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000276:immunology*; D008895:Milk, Human / Q000276:immunology*; D008970:Molecular Weight
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D007071:Immunoglobulin A, Secretory