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

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Abstract
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.

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