AIMS:CAWS, Candida albicans water-soluble fraction, is an extracellular mannoprotein produced by C. albicans NBRC1385. It is a ligand of dectin-2, the C-type lectin receptor for innate immunity, and has strong potency for induction of vasculitis in DBA/2 mice. The structure of this mannoprotein is known to be modulated by the culture conditions. To clarify the structure required for vasculitis, CAWSs were prepared in the two culture conditions with or without pH control, and biological properties were compared. METHODS:CAWSs prepared by the standard protocol and pH controlled at 7.0 were designated as CAWS and CAWS727, respectively. The antigenicity was detected by the anti-Candida mannan IgG. These chemical structures were assessed by nuclear magnetic resonance analysis and the lectin array system. The in vitro activity of CAWSs was tested by tumor necrosis factor-α (TNF-α) induction using bone marrow-derived dendritic cells and spleen cell cultures. RESULTS:The antigenicity of CAWS727 was similar to CAWS but the nuclear magnetic resonance analysis showed a higher ratio of β-mannosyl linkages were detected in CAWS727. The lectin array showed relative affinities of CAWS727 to α-mannosyl specific lectins were weaker than those of CAWS. CAWS induced severe vasculitis in DBA/2 mice while CAWS727 did not. CAWS significantly induced TNF-α but CAWS727 did slightly. In addition, CAWS-induced TNF-α production was inhibited by mixing with CAWS727 in a concentration dependent manner. CONCLUSION:The α-mannosyl linkages of Candida mannan is a key molecule for the immunotoxicity. CAWS727, which conatins β-mannosyl linkages, competitively bound to lectin receptors, and resulted in reductions in the inflammatory response.

译文

目的:CAWS,白色念珠菌水溶性级分,是白色念珠菌NBRC1385产生的细胞外甘露糖蛋白。它是先天性免疫的C型凝集素受体dectin-2的配体,对DBA / 2小鼠的血管炎具有很强的诱导作用。已知该甘露糖蛋白的结构受培养条件调节。为了阐明血管炎所需的结构,在两种培养条件下(有或没有pH值控制)制备了CAWS,并比较了生物学特性。
方法:将标准方案制备的且pH控制在7.0的CAWS分别命名为CAWS和CAWS727。通过抗Candida甘露聚糖IgG检测抗原性。这些化学结构通过核磁共振分析和凝集素阵列系统进行评估。使用骨髓来源的树突状细胞和脾细胞培养物,通过肿瘤坏死因子-α(TNF-α)诱导来测试CAWS的体外活性。
结果:CAWS727的抗原性与CAWS相似,但核磁共振分析表明在CAWS727中检测到较高比例的β-甘露糖基键。凝集素阵列显示CAWS727对α-甘露糖基特异性凝集素的相对亲和力弱于CAWS。 CAWS诱发了DBA / 2小鼠的严重血管炎,而CAWS727并未诱发。 CAWS显着诱导TNF-α,但CAWS727略有诱导。另外,通过以浓度依赖性方式与CAWS727混合来抑制CAWS诱导的TNF-α的产生。
结论:甘露假丝酵母的α-甘露糖基键是免疫毒性的关键分子。 CAWS727具有β-甘露糖基键,与凝集素受体竞争性结合,并导致炎症反应降低。

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