Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan.
Immunopharmacol Immunotoxicol. 2011 Jun;33(2):302-8. doi: 10.3109/08923973.2010.503707. Epub 2010 Jul 30.
We have previously obtained and elucidated the precise structure of a highly branched 1,3-β-D-glucan (with 6-monoglucopyranosyl side chains), Aureobasidium pullulans-fermented β-D-glucan (AP-FBG), from the fungus A. pullulans. However, the mechanism(s) of the effects of AP-FBG on in vitro mouse primary cells have not been analyzed in detail. Herein, we report that the induction of cytokines by AP-FBG was dependent on the existence of a granulocyte macrophage colony-stimulating factor (GM-CSF); this is similar way to be a typical 1,3-β-D-glucan from Sparassis crispa (SCG), which is a 1,3-β-D-glucopyranosyl backbone with single 1,6-β-D-glucopyranosyl side branching units every three residues. In other words, the production of cytokines in DBA/2-mouse-derived splenocytes by AP-FBG was completely hampered by an anti-GM-CSF neutralizing monoclonal antibody. Furthermore, the addition of exogenous GM-CSF to C57BL/6-derived splenocytes, which are less sensitive to AP-FBG, induced the production of cytokines by AP-FBG. Therefore, GM-CSF is indispensable for the induction of cytokines by AP-FBG in mouse-derived splenocytes. This finding has provided a new insight into our understanding of the actions of β-D-glucan but will also aid in the design and development of more effective β-D-glucan agents.
我们之前已经获得并阐明了一种高度分支的 1,3-β-D-葡聚糖(具有 6-单葡萄糖吡喃基侧链),即出芽短梗霉发酵的β-D-葡聚糖(AP-FBG),来自真菌出芽短梗霉。然而,AP-FBG 对体外小鼠原代细胞的影响机制尚未详细分析。在此,我们报告称,AP-FBG 诱导细胞因子的产生依赖于粒细胞-巨噬细胞集落刺激因子(GM-CSF)的存在;这与来自裂褶菌(SCG)的典型 1,3-β-D-葡聚糖相似,SCG 是一种具有单 1,6-β-D-葡萄糖吡喃基侧分支单元的 1,3-β-D-葡聚糖主链,每三个残基存在一个。换句话说,AP-FBG 完全抑制了抗 GM-CSF 中和单克隆抗体对 DBA/2 小鼠来源的脾细胞中细胞因子的产生。此外,外源性 GM-CSF 添加到对 AP-FBG 敏感性较低的 C57BL/6 衍生的脾细胞中,可诱导 AP-FBG 产生细胞因子。因此,GM-CSF 对于 AP-FBG 在小鼠来源的脾细胞中诱导细胞因子的产生是不可或缺的。这一发现为我们理解β-D-葡聚糖的作用提供了新的见解,也将有助于设计和开发更有效的β-D-葡聚糖制剂。