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用单克隆抗体对霍乱弧菌O:1脂多糖、O侧链和核心进行免疫学特性分析。

Immunological characterization of Vibrio cholerae O:1 lipopolysaccharide, O-side chain, and core with monoclonal antibodies.

作者信息

Gustafsson B, Holme T

出版信息

Infect Immun. 1985 Aug;49(2):275-80. doi: 10.1128/iai.49.2.275-280.1985.

Abstract

Lipopolysaccharide (LPS) was extracted from Vibrio cholerae O:1 strains of the serotypes Ogawa, Inaba, and Hikojima and delipidated by mild-acid hydrolysis. Two polysaccharide fragments with the molecular weights of approximately 9,000 and 900, respectively, were isolated by gel permeation chromatography. The LPS preparations and the polysaccharide fragments were studied in enzyme-linked immunosorbent assay inhibition, rocket immunoelectrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroblotting with monoclonal antibodies directed against the group-specific antigen A, the type-specific antigens B (Ogawa) and C (Inaba), and the core region. Antigen A was demonstrated in all LPS preparations and all 9,000-molecular-weight fragments tested. The type-specific antigens B and C were demonstrated in LPSs and 9,000-molecular-weight fragments from Ogawa and Inaba, respectively. Furthermore, antigens B and C were both demonstrated in LPSs and 9,000-molecular-weight fragments from two of four Hikojima strains tested. Core antigen was demonstrated in the LPS and in the 9,000- and 900-molecular-weight fragments. The results indicate that the 9,000-molecular-weight fragment represents the complete polysaccharide chain, including group- and type-specific antigens as well as core antigens, whereas the 900-molecular-weight fragment constitutes the main part of the core region.

摘要

从霍乱弧菌O:1小川型、稻叶型和彦岛型菌株中提取脂多糖(LPS),并通过温和酸水解进行脱脂处理。通过凝胶渗透色谱法分离出两种分子量分别约为9000和900的多糖片段。利用针对群特异性抗原A、型特异性抗原B(小川型)和C(稻叶型)以及核心区域的单克隆抗体,对LPS制剂和多糖片段进行了酶联免疫吸附测定抑制试验、火箭免疫电泳以及十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,随后进行电印迹分析。在所有测试的LPS制剂和所有分子量为9000的片段中均检测到抗原A。在小川型和稻叶型的LPS及分子量为9000的片段中分别检测到型特异性抗原B和C。此外,在测试的4株彦岛型菌株中的2株的LPS及分子量为9000的片段中均检测到抗原B和C。在LPS以及分子量为9000和900的片段中均检测到核心抗原。结果表明,分子量为9000的片段代表完整的多糖链,包括群特异性和型特异性抗原以及核心抗原,而分子量为900的片段构成核心区域的主要部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7da6/262010/fc94d8ede674/iai00113-0029-a.jpg

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