Preparation, characterization, and immunogenicity of Haemophilus influenzae type b polysaccharide-protein conjugates.
Preparation, characterization, and immunogenicity of Haemophilus influenzae type b polysaccharide-protein conjugates.
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DOI：
10.1084/jem.152.2.361
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发表时间：
1980-08-01
期刊：
影响因子：
--
通讯作者：
Robbins JB
中科院分区：
文献类型：
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作者：
Schneerson R;Barrera O;Sutton A;Robbins JB
提出了一种将 b 型流感嗜血杆菌荚膜多糖 (HIB Ps) 与多种蛋白质共价结合的方法。该方法是有效的并且依赖于使用己二酸二酰肼作为荚膜多糖和载体蛋白之间的间隔物。与纯化的 HIB Ps 在小鼠和兔子中的免疫原性差相反，HIB Ps-蛋白缀合物诱导血清中具有杀菌活性的抗 b 型抗体，其水平在皮下注射盐溶液中的低剂量时显示出对人类具有保护作用。小鼠体内的抗体反应与缀合物的剂量有关，随着注射次数的增加而增加，并且可以通过先前注射载体蛋白来引发。 HIB Ps-蛋白缀合物在三种不同的小鼠品系中具有免疫原性。用同源多糖或3型肺炎球菌多糖制备的HIB Ps杂交体未能在小鼠中诱导抗体，这表明了载体分子对于增强HIB Ps-蛋白缀合物免疫原性的重要性。注射弗氏佐剂乳化的HIB Ps-蛋白缀合物的兔子产生高水平的血清抗b型抗体，该抗体诱导对b型流感嗜血杆菌生物体的杀菌作用。据推测，多糖蛋白缀合物的 HIB Ps 成分已转化为胸腺依赖性免疫原。该方法可用于制备具有 HIB Ps 和其他考虑供人类使用的多糖的蛋白质-多糖缀合物。
A method is presented for covalently bonding Haemophilus influenzae type b capsular polysaccharide (HIB Ps) to several proteins. The method is efficient and relies upon the use of adipic dihydrazide as a spacer between the capsular polysaccharide and the carrier protein. In contrast to the poor immunogenicity of the purified HIB Ps in mice and rabbits, the HIB Ps-protein conjugates induced serum anti-type b antibodies having bactericidal activity at levels shown to be protective in humans when low doses were injected subcutaneously in a saline solution. The antibody response in mice was related to the dose of the conjugates, increased with the number of injections, and could be primed by the previous injection of the carrier protein. The HIB Ps- protein conjugates were immunogenic in three different mouse strains. The importance of the carrier molecule for the enhanced immunogenicity of the HIB Ps-protein conjugates was shown by the failure of HIB Ps hybrids prepared with either the homologous polysaccharide or pneumococcus type 3 polysaccharide to induce antibodie in mice. Rabbits injected with the HIB Ps-protein conjugates emulsified in Freund's adjuvant produced high levels of serum anti-type b antibodies which induced a bactericidal effect upon H. influenzae type b organisms. It is proposed that the HIB Ps component of the polysaccharide protein conjugates has been converted to a thymic-dependent immunogen. This method may be used to prepare protein-polysaccharide conjugates with HIB Ps and other polysaccharides to be considered for human use.