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Immunogenicity of Vibrio vulnificus capsular polysaccharides and polysaccharide-protein conjugates.

机译:创伤弧菌荚膜多糖和多糖-蛋白质结合物的免疫原性。

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摘要

Opaque colony morphology has been correlated to Vibrio vulnificus virulence. However, the number of capsular serotypes expressed by virulent isolates is unknown. In an effort to produce anticapsule sera, capsular polysaccharide (CPS) from three opaque V. vulnificus strains was purified and characterized. Purified CPSs were acidic and contained considerable amounts of hexosamine and trace quantities of protein and nucleic acid. CPS purified from strain C7184 was poorly immunogenic for rabbits and mice, since repeated injection produced little detectable anticapsular antibody. To improve immunogenicity, CPS-protein conjugates were prepared from adipic acid hydrazide derivatives of CPS purified from each strain and carbodiimide as a coupling reagent. The immunogenicity of C7184 CPS was enhanced by conjugation to keyhole limpet hemocyanin, since injection into mice elicited production of anticapsular antibodies, the level of which was dependent on the dose and time since initial immunization. Injection of rabbits with CPS-protein conjugates also produced anticapsular antibodies. Staphylococcus aureus cells armed with each of the three anticapsular antibodies coagglutinated only the homologous opaque strain, indicating the existence of at least three capsular types. Further screening of 32 opaque and translucent V. vulnificus isolates revealed only three cross-reacting strains. These results suggest the presence of numerous V. vulnificus capsular types.
机译:不透明的菌落形态已与创伤弧菌毒力相关。但是,由毒力分离株表达的荚膜血清型的数目是未知的。为了产生抗胶囊血清,纯化和表征了来自三个不透明的V. vulnificus菌株的荚膜多糖(CPS)。纯化的CPS是酸性的,含有大量的六胺和痕量的蛋白质和核酸。从C7184株纯化的CPS对兔和小鼠的免疫原性很差,因为重复注射几乎没有产生可检测的抗荚膜抗体。为了提高免疫原性,由从各菌株纯化的CPS的己二酸酰肼衍生物和碳二亚胺作为偶联剂制备CPS-蛋白缀合物。 C7184 CPS的免疫原性可通过与匙孔血蓝蛋白结合而增强,因为向小鼠注射可产生抗荚膜抗体,抗荚膜抗体的水平取决于首次免疫后的剂量和时间。向兔子注射CPS-蛋白结合物也产生抗荚膜抗体。装备有三种抗荚膜抗体的金黄色葡萄球菌细胞仅凝集同源的不透明菌株,表明存在至少三种荚膜类型。进一步筛选了32个不透明和半透明的V. vulnificus分离株,发现只有三个交叉反应菌株。这些结果表明存在大量的V. vulnificus荚膜类型。

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