PURIFICATION AND IMMUNOGENICITY OF GENETICALLY OBTAINED PNEUMOLYSIN TOXOIDS AND THEIR CONJUGATION TO STREPTOCOCCUS-PNEUMONIAE TYPE-19F POLYSACCHARIDE
PURIFICATION AND IMMUNOGENICITY OF GENETICALLY OBTAINED PNEUMOLYSIN TOXOIDS AND THEIR CONJUGATION TO STREPTOCOCCUS-PNEUMONIAE TYPE-19F POLYSACCHARIDE
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DOI：
10.1128/iai.59.7.2297-2304.1991
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发表时间：
1991-07-01
影响因子：
3.1
通讯作者：
BOULNOIS, GJ
中科院分区：
文献类型：
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作者：
PATON, JC;LOCK, RA;BOULNOIS, GJ
作为一项正在进行的关于改进人类肺炎链球菌疫苗的研究的一部分，将两个已定义的肺炎溶血素(PL)类毒素(肺炎溶菌体)的基因插入到大肠杆菌高效表达载体pKK233-2中，并对其进行了纯化。用PD-A、PD-B或从肺炎链球菌提纯的天然PL免疫的组小鼠，然后用强毒肺炎球菌鼻腔或腹膜内攻击。所有免疫组的小鼠存活时间明显长于假免疫对照组。作为保护性免疫原，两种溶血素均比磷脂更有效。将肺炎球菌19F衣壳多糖(19F PS)与肺炎球菌PD-B共价偶联，测定其蛋白和PS部分在小鼠体内的免疫原性。获得了显著的抗PL效价，19F PS部分的免疫原性比未结合PS显著增强。结合似乎也已将19F PS转化为能够诱导增强效应的抗原。这些结果支持这样的观点，即以PS为基础的人类抗肺炎球菌疫苗的效力可以通过以共价PS-PS结合物的形式补充类肺炎蛋白来提高。
As part of an ongoing study concerned with improving human vaccines against Streptococcus pneumoniae, the genes for two defined pneumolysin (PL) toxoids (pneumolysoids), Pd-A (PL with a Cys-->Gly substitution at amino acid 428) and Pd-B (PL with a Trp-->Phe substitution at position 433), were inserted into the high-expression vector pKK233-2 in Escherichia coli and the pneumolysoids were purified. Groups of mice which had been immunized with either Pd-A, Pd-B, or native PL purified from S. pneumoniae were then challenged either intranasally or intraperitoneally with virulent pneumococci. Mice in all immunized groups survived significantly longer than sham-immunized controls. Both pneumolysoids were more effective than PL as protective immunogens. Pneumolysoid Pd-B was conjugated covalently with pneumococcal type 19F capsular polysaccharide (19F PS), and the immunogenicities of both the protein and the PS moieties of the conjugate in mice were determined. Significant anti-PL titers were obtained, and the immunogenicity of the 19F PS moiety was markedly enhanced compared with that of unconjugated PS. Conjugation also appears to have converted the 19F PS into an antigen capable of inducing a booster effect. These results support the notion that the efficacy of human, PS-based antipneumococcal vaccines might be improved by supplementation with pneumolysoid in the form of a covalent pneumolysoid-PS conjugate.