Two known Clostridiodes (Clostridium) difficile surface antigens, a lipoteichoic acid (LTA) and a polysaccharide (PS-II) were isolated and purified in order to prepare glycoconjugate vaccines to the carrier protein human serum albumin utilising a reductive amination strategy. Mice and rabbits were immunized with a prime and two boost strategy and the resulting sera were examined for their ability to recognise the purified homologous antigens and subsequently killed whole cells of C. difficile strains and other Clostridia species. Immunisation derived antisera from rabbits and mice, recognised all strains of C. difficile vegetative cells examined, with generally similar titers from animals that received the LTA or the PS-II conjugates. Sera raised to the LTA conjugates were able to recognise other Clostridia species C. butyricum, C. bifermentans and C. subterminale whereas sera raised to the PS-II conjugates were not. These LTA and PS-II sera recognised live cells in an immunofluorescence assay and were also able to recognise the spore form of the bacterium. This study has confirmed that the LTA and PS-II polysaccharides are both highly conserved surface polymers of C. difficile that are easily accessible to the immune system and as such may have potential as vaccine antigens or as targets for therapeutics to combat C. difficile infection.

分离和纯化两种已知的艰难梭菌 (Clostridium)表面抗原脂磷壁酸 (LTA) 和多糖 (PS-II)，以利用还原胺化策略制备针对载体蛋白人血清白蛋白的糖结合疫苗。用初免和两次加强策略免疫小鼠和兔子，并检查所得血清识别纯化的同源抗原并随后杀死艰难梭菌菌株和其他梭菌属物种的全细胞的能力。来自兔和小鼠的免疫衍生抗血清可识别所有艰难梭菌菌株检查营养细胞，与接受 LTA 或 PS-II 缀合物的动物的滴度大致相似。用 LTA 偶联物培养的血清能够识别其他梭菌种丁酸梭菌、双发酵梭菌和近末端梭菌，而用 PS-II 偶联物培养的血清则不能。这些 LTA 和 PS-II 血清在免疫荧光测定中识别活细胞，并且还能够识别细菌的孢子形式。该研究证实，LTA 和 PS-II 多糖都是艰难梭菌的高度保守的表面聚合物，免疫系统很容易接触到，因此可能具有作为疫苗抗原或作为对抗艰难梭菌的治疗靶点的潜力感染。