Vaccine Program, Human Health Therapeutics Portfolio, National Research Council, Ottawa, ON, K1A 0R6, Canada,
Glycoconj J. 2013 Dec;30(9):843-55. doi: 10.1007/s10719-013-9489-3. Epub 2013 Aug 23.
A lipoteichoic acid has recently been shown to be conserved in the majority of strains from Clostridium difficile and as such is being considered as a possible vaccine antigen. In this study we examine the candidacy of the conserved lipoteichoic acid by demonstrating that it is possible to elicit antibodies against C. difficile strains following immunisation of rabbits and mice with glycoconjugates elaborating the conserved lipoteichoic acid antigen. The present study describes a conjugation strategy that utilises an amino functionality, present at approximately 33 % substitution of the N-acetyl-glucosamine residues within the LTA polymer repeating unit, as the attachment point for conjugation. A maleimide-thiol linker strategy with the maleimide linker on the carboxyl residues of the carrier protein and the thiol linker on the carbohydrate was employed. Immunisation derived antisera from rabbits and mice, recognised all strains of C. difficile vegetative cells examined, despite an immune response to the linkers also being observed. These sera recognised live cells in an immunofluorescence assay and were also able to recognise the spore form of the bacterium. This study has illustrated that the LTA polymer is a highly conserved surface polymer of C. difficile that is easily accessible to the immune system and as such merits consideration as a vaccine antigen to combat C. difficile infection.
最近已经证明，艰难梭菌的大多数菌株都含有脂磷壁酸，因此它被认为是一种可能的疫苗抗原。在这项研究中，我们通过证明在使用糖缀合物免疫兔子和小鼠后，有可能针对艰难梭菌菌株产生抗体，从而检查了保守的脂磷壁酸的候选性。本研究描述了一种利用存在于 LTA 聚合物重复单元中的 N-乙酰葡萄糖胺残基的约 33%取代的氨基功能作为缀合点的缀合策略。使用带有载体蛋白羧基的马来酰亚胺-硫醇接头的马来酰亚胺接头和碳水化合物上的硫醇接头的马来酰亚胺-硫醇接头策略。从兔子和小鼠中获得的免疫血清识别所有检查的艰难梭菌营养细胞菌株，尽管也观察到对接头的免疫反应。这些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。这项研究表明，LTA 聚合物是艰难梭菌的一种高度保守的表面聚合物，很容易被免疫系统识别，因此值得考虑作为对抗艰难梭菌感染的疫苗抗原。
