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概要

C. difficile毒素 Bの受容体結合ドメインの生化学的および免疫学的特性

ジュイシン・ファン、チアウェイ・ウー、シュウペイ・リアン、クアンナン・シャオ、チーシャン・レン、イアン・ユーシン・リン、ペレ・チョン

Clostridium difficile (Cd) is an emerging nosocomial pathogen responsible for antibiotic-associated pseudo-membranous colitis and diarrhea in hospital acquired infections. Clostridial toxins A (TcdA) and B (TcdB) which specifically bind to unknown glycoprotein(s) on the surface of epithelial cells disrupt the intestinal barrier and ultimately lead to acute inflammation and diarrhea . There is still debate as to whether the receptor binding domains (RBD) of toxins can individually elicit protection in the hamster challenge model. In this study, a TcdB RBD which was derived from C.difficile strain VPI10463 with >95% amino acid sequence identity to hyper-virulent strain BI/NAP1/027 was designed and expressed in Escherichia coli. Recombinant RBD (rRBD) was purified, characterized biologically and immunologically and found to have the following properties: (a) capable of binding to the cell surface of both Vero and Caco-2 cells and entering into the cytosol; (b) devoid of hemagglutinin activity (HA); (c) the ability to up-regulate cell surface markers expressions and cytokines secretions from dendritic cells; (d) eliciting anti-TcdB neutralizing antibody responses that could weakly cross-neutralize TcdA in the absence of adjuvant; (e) and inducing weak protection against a lethal dose of Cd spores in the hamster challenge model. Therefore, rRBD shows potential as an immunogen to be included in the development of vaccines against Clostridium difficile-associated diseases.

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