Haemaglutination of Shigella dysenteriae subunit pili protein with anti-haemaglutination of S. dysenteriae subunit pili protein as a molecule adhesion in mouse enterocytes

摘要

Shigellosis is a global human health problem. Unfortunately, there has not been any effective vaccine available worldwide. Molecule adhesion of bacteria can be used as a component of vaccine as it facilitates attachment to a cell surface structure in which the adhesion molecule is located in the pili. The purpose of this study was to clarify whether the protein subunit pili Shigella dysenteriae has a molecule adhesin. The purification of pili was done by isolating them using a pilus bacterium cutter. Purification of protein subunit pili resulting from SDS-PAGE was obtained by an electro-elution method. Adherent assays for mice enterocyte were used by conducting in a dose dependent manner and by doing an immuno-cytochemistry. The purified pili proteins with MW 49.8 kDa showed a haemagglutinin towards mouse erythrocytes. The pili proteins with MW 7.9 kDa showed an anti-haemaggutinin if added to a haemaggutinin pili proteins with MW 49.8 kDa and can prevent haemagglutination. Furthermore, pretreatment of the enterocytes purified with MW 49.8 and 7.9 kDa pili proteins, the adherence of S. dysenteriae to mouse enterocytes was inhibited. Immuno-cytochemistry showed that haemaggutinin protein with MW 49.8 kDa and anti-haemagglutinin protein with MW 7.9 kDa adhered to mouse erythrocyte. These results suggest that haemaggutinin protein with MW 49.8 kDa and anti-haemaggutinin protein with MW 7.9 kDa of S. dysenteriae pili proteins are adhesive proteins involved in the S. dysenteriae initial adherence mechanisms for the enterocytes.