The ability to adhere via colonization factors to specific receptors located on the intestinal mucosa is a key virulence factor in enterotoxigenic Escherichia coli (ETEC) pathogenesis. Here, the potential glycosphingolipid receptors of the novel human ETEC colonization factor CS30 were examined by binding of CS30-expressing bacteria to glycosphingolipids on thin-layer chromatograms. We thereby found a highly specific binding of CS30-expressing bacteria to a fast-migrating acid glycosphingolipid of human and porcine small intestine, while no binding was obtained with a mutant ETEC strain unable to express CS30 fimbriae. The CS30 binding glycosphingolipid from human small intestine was isolated and characterized by mass spectrometry as sulfatide (SO3-3Galβ1Cer). Comparative binding studies using sulfatides with different ceramide compositions gave a preferential binding of CS30 to sulfatide with d18:1-h24:0 ceramide. This ceramide species of sulfatide was also isolated from human small intestine and characterized by mass spectrometry and antibody binding. These studies implicate sulfatide as candidate receptor for mediating attachment of CS30-fimbriated ETEC to human and porcine small intestinal cells. Our findings may be a basis for designing receptor saccharide analogues for inhibition of the intestinal adhesion of CS30-expressing E. coli.

• Klíčová slova
• Enterotoxigenic E. coli, carbohydrate binding, colonization factor CS30, glycosphingolipid characterization, microbial adhesion, sulfatide,
• MeSH
• bakteriální adheze * MeSH
• ceramidy analýza   MeSH
• enterotoxigenní Escherichia coli metabolismus   MeSH
• faktory virulence metabolismus   MeSH
• glykosfingolipidy metabolismus   MeSH
• lidé MeSH
• prasata MeSH
• proteiny fimbrií genetika   metabolismus   MeSH
• proteiny z Escherichia coli metabolismus   MeSH
• střevní sliznice cytologie   mikrobiologie   MeSH
• sulfoglykosfingolipidy metabolismus   MeSH
• tenké střevo cytologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ceramidy MeSH
• colonization factor antigens MeSH Prohlížeč
• faktory virulence MeSH
• glykosfingolipidy MeSH
• proteiny fimbrií MeSH
• proteiny z Escherichia coli MeSH
• sulfoglykosfingolipidy MeSH

Primers were designed and prepared and conditions were determined for PCR detection and differentiation of enterotoxigenic E. coli bacterial strains isolated from diarrheic pigs. Primers K88/1 and K88/2 are 25 bp oligomers that correspond to a region of genes encoding one of serological variants of the K88 antigen (K88ab(1), K88ab(2), K88ac or K88ad). A positive result of PCR is an amplificate of 792 bp in size for K88ab and K88ad variant or 786 bp for K88ac variant. The individual serological variants of genes of the K88 antigen could be differentiated by cutting the obtained PCR amplificates by restriction endonucleases. The PCR analysis of 674 E. coli strains isolated from diarrheic pigs showed that 184 strains were K88 positive. By using restriction endonucleases the K88-positive strains were in 4 cases classified as K88ab variant, 180 as K88ac variant and none contained gene for the K88ad variant. Ninety-five % coincidence with serological examination using K88ab, K88ac and K88ad specific antibodies was shown.

• MeSH
• antigeny bakteriální analýza   klasifikace   genetika   MeSH
• DNA primery MeSH
• Escherichia coli klasifikace   genetika   izolace a purifikace   MeSH
• genetická variace MeSH
• infekce vyvolané Escherichia coli mikrobiologie   veterinární   MeSH
• nemoci prasat mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• prasata MeSH
• proteiny fimbrií analýza   klasifikace   genetika   MeSH
• proteiny z Escherichia coli analýza   klasifikace   genetika   MeSH
• průjem mikrobiologie   veterinární   MeSH
• restrikční enzymy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika MeSH
• Názvy látek
• antigeny bakteriální MeSH
• DNA primery MeSH
• K88 antigen, E coli MeSH Prohlížeč
• proteiny fimbrií MeSH
• proteiny z Escherichia coli MeSH
• restrikční enzymy MeSH

One-hundred sixty Escherichia coli isolates obtained from piglets with diarrhea from different parts of Slovakia were examined for the presence of genes coding for F4, F5, F6 and F41 fimbrial adhesins, and hemolytic activity. According to polymerase chain reaction tests 74 (46%) E. coli isolates were positive for primers that detected genes coding for fimbrial adhesins. Of these 74 isolates, 64 were positive for genes encoding for F4+, four for F5+, five for F6+, and one for both F41+ and F5+ adhesins.

• MeSH
• adheziny Escherichia coli genetika   MeSH
• antigeny bakteriální genetika   MeSH
• bakteriální adheziny genetika   MeSH
• bakteriální toxiny biosyntéza   MeSH
• DNA bakterií analýza   genetika   MeSH
• Escherichia coli genetika   izolace a purifikace   MeSH
• genotyp MeSH
• hemolyziny biosyntéza   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   veterinární   MeSH
• nemoci prasat mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• prasata mikrobiologie   MeSH
• proteiny z Escherichia coli genetika   MeSH
• průjem mikrobiologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika MeSH
• Názvy látek
• adheziny Escherichia coli MeSH
• antigeny bakteriální MeSH
• bakteriální adheziny MeSH
• bakteriální toxiny MeSH
• DNA bakterií MeSH
• F41 antigen, E coli MeSH Prohlížeč
• hemolyziny MeSH
• proteiny z Escherichia coli MeSH