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首页> 外文期刊>Infection and immunity >Mapping of Candida albicans oligomannosidic epitopes by using monoclonal antibodies.
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Mapping of Candida albicans oligomannosidic epitopes by using monoclonal antibodies.

机译：通过使用单克隆抗体对白色念珠菌寡甘露糖苷表位进行定位。

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摘要

Six monoclonal antibodies (MAbs) from various laboratory sources (EB-CA1, EB-CA2, H5, AF1, C6, and 5B2), reacting with the polysaccharidic moieties of Candida albicans mannoproteins, were used for epitope mapping by an enzyme-linked immunosorbent assay (ELISA) with neoglycolipids and by Western blotting (immunoblotting) of a C. albicans germ tube extract. The ELISA involved neoglycolipids constructed from three families of oligomannosides released by sequential depolymerization of C. albicans phosphopeptidomannan by acid hydrolysis (NGLH), beta-elimination (NGLO), and acetolysis (NGLA). All of the MAbs exhibited low reactivities against NGLO. MAbs EB-CA1, EB-CA2, and H5 reacted mainly against NGLA, and MAbs C6 and AF1 recognized mainly NGLH, whereas MAb 5B2 reacted with both families of neoantigens. When this method was compared with Western blotting, strong reactivity to NGLA was associated with the presence of epitopes shared by high-molecular-weight mannoproteins, whereas strong reactivity to NGLH was associated with a reactivity to a family of 14- to 18-kDa antigens. The reactivity of MAb 5B2 was associated with both high-molecular-weight mannoproteins and the 14- to 18-kDa antigens. In relation to the present knowledge about the structure of the C. albicans phosphopeptidomannan oligomannosidic repertoire, these results provide preliminary data concerning the molecular basis of the recognition of mannopyranosyl sequences by MAbs and their distribution among C. albicans mannoproteins.
机译：与多种白色念珠菌甘露糖蛋白的多糖部分反应的六种单克隆抗体（MAb）来自多种实验室来源（EB-CA1，EB-CA2，H5，AF1，C6和5B2），通过酶联免疫吸附剂用于表位作图新糖脂和白色念珠菌胚芽管提取物的蛋白质印迹法（免疫印迹）进行免疫分析（ELISA）。 ELISA涉及由三个家族的寡甘露糖苷构成的新糖脂，该寡糖甘露糖苷是通过酸水解（NGLH），β-消除（NGLO）和乙酰分解（NGLA）依次分离白色念珠菌磷酸肽甘露聚糖而释放的。所有的单克隆抗体都表现出对NGLO的低反应性。单克隆抗体EB-CA1，EB-CA2和H5主要针对NGLA反应，单克隆抗体C6和AF1主要识别NGLH，而单克隆抗体5B2与两个新抗原家族均发生反应。当将该方法与蛋白质印迹法进行比较时，对NGLA的强反应性与高分子量甘露糖蛋白共有的表位的存在有关，而对NGLH的强反应性与对14至18 kDa抗原家族的反应性有关。 MAb 5B2的反应性与高分子量甘露糖蛋白和14至18 kDa抗原相关。关于白念珠菌磷酸肽甘露聚糖寡甘露糖苷库的结构的现有知识，这些结果提供了关于MAb识别甘露吡喃糖基序列的分子基础及其在白念珠菌甘露糖蛋白中的分布的初步数据。

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