Glycosphingolipids as Putative Receptor for Staphylococcal Enterotoxin-B in Cultured Human Kidney Cells.
Annual rept. 1 Aug 95-31 Jul 96,
JOHNS HOPKINS UNIV BALTIMORE MD
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Staphylococcal enterotoxin-B SEB is a common enterotoxin that can cause diarrhea and death in man. Since, we found that digalactosylceramide is most probably a putative receptor for SEB in cultured human kidney proximal tubular cells, it was used to develop a specific and sensitive assay for SEB enzyme linked receptor-based immunodot in human fluids, plasma and urine. Digalactosylceramide was immobilized on a polyvinyledene difluoride membrane and the membrane was subsequently incubated with primary and secondary alkaline phosphatase conjugated antibodies. A positive reaction was discerned as a blue spot. As little as 1 ugml SEB could be detected in the assay. SEB did not bind to structurally related glycosphingolipids, indicating specificity. This assay was specific for SEB, but not for SEA and TSST-I. Further work is in progress to study the application of this method in the field. Our findings will be of potential value for the food industry, and to determine toxemia in our soldiers.