Oral infection with enteropathogenic Escherichia coli triggers immune response and intestinal histological alterations in mice selected for their minimal acute inflammatory responses.

Enteropathogenic Escherichia coli (EPEC), a leading cause of infant diarrhea, is an important public health problem in Brazil and other developing countries. In vitro assays of bacterial adhesion to cultured cells are important tools for studying bacterial pathogenicity but do not reproduce all the events that occur in natural infections. In this study, the effects of oral infection with EPEC on mice selected for their minimal acute inflammatory response (AIR min) were evaluated. Mice were orally infected with EPEC and variations in body weight, bacterial shedding and antibody production observed. The infected animals developed seric and secretory anti-EPEC antibodies; however, neither mortality nor diarrhea was observed. Light microscopy of their intestines demonstrated histological modifications that were not present in controls. However, electron microscopy did not show bacteria attached to the intestinal epithelia to form attaching and effacing lesions, characteristic of EPEC in humans. The bacteria were detected in Peyer's patches and intestinal contents up to 5 hr post-infection. When human anti-EPEC secretory immunoglobulin A or avian immunoglobulin Y antibodies were administered to infected animals, they developed minor histological alterations compared with non-treated animals. In summary, it was found that EPEC triggers immune responses and intestinal histological alterations but does not produce evidence of diarrheal disease in mice infected by the oral route. This study of EPEC experimental infection provides a better understanding of the effects of antibodies on bacterial infections and may provide a suitable model for the design and testing of immunobiological products for active or passive immunization.

Seric and secretory antibodies reactive to alpha, beta and gamma intimins of Escherichia coli in healthy Brazilian adults.

Intimin is essential for attaching and effacing lesions by pathogens such as enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC), and the antigenic polymorphism of intimin determines distinct subtypes. Our aim was to investigate the presence of immunoglobulin G (IgG) and IgA antibodies reactive to alpha, beta and gamma intimins in serum and colostrum from healthy Brazilian adults. We found seric IgG and secretory IgA antibodies reactive to conserved and variable regions of alpha, beta and gamma intimins and a positive correlation between the concentrations of these antibodies in both serum and colostrum that suggested cross reactivity among anti-intimin antibodies, as was confirmed by immunoblotting and absorption. The concentrations of anti-conserved region antibodies were higher than those of variable region antibodies. The presence of antibodies reactive to EHEC antigens could result from contact with EPEC or with other bacteria of the environment even though this bacterium is not frequent in Brazil, and suggests possible protection against EHEC.

Early acquisition of serum and saliva antibodies reactive to enteropathogenic Escherichia coli virulence-associated proteins by infants living in an endemic area.

Enteropathogenic Escherichia coli (EPEC) is the most common etiological agent of acute diarrhea among infants living in poor social conditions in Brazil and other developing countries. This infection is rare in breast-fed infants, as well as in children older than 2 years. Over the past few years, our group has attempted to identify antibodies to EPEC virulence proteins in human milk and to establish the in vitro protective role of these antibodies. In the present study, we report the identification of antibodies to EPEC virulence proteins in sera and saliva from children of different ages, living in slums in the city of São Paulo, Brazil. Using EPEC and bacterial constructs (pET) for immunoblotting (IB) analysis, antibodies reacting to the main adhesins (intimin, bundle-forming pilli) and cell-signaling proteins (EPEC secreted proteins - Esp A, Esp B) were detected in sera from adults and children older than 1 year. Almost all children older than 1 year presented recognition patterns similar to those of adults in IB assays for serum IgG and secretory IgA antibodies, using EPEC outer membrane and other antigenic preparations. As previously observed for human milk, all samples from adults and older children recognized the 94 kDa molecular weight adhesin intimin strongly. In most children, previous EPEC symptomatic diarrhea could not be confirmed; however, almost all of them have presented one or more diarrhea episodes during their lifetime. These results suggest that reduction of EPEC infection frequency after 2 years of age may be associated with the development of anti-EPEC antibody repertoires.