Dissociation of toll-like receptor 2-mediated innate immune response to zymosan by organic solvent-treatment without loss of dectin-1 reactivity
Dissociation of toll-like receptor 2-mediated innate immune response to zymosan by organic solvent-treatment without loss of dectin-1 reactivity
复制标题
DOI：
10.1248/bpb.31.13
复制
发表时间：
2008-01-01
影响因子：
2
通讯作者：
Ohno, Naohito
中科院分区：
文献类型：
--
作者：
Ikeda, Yoshihiko;Adachi, Yoshiyuki;Ohno, Naohito
酵母多糖通过天然免疫系统激活吞噬细胞，引起动物的炎症反应。由于酵母多糖制剂中所含活性物质的复杂性，需要简化实际负责天然免疫识别的活性部分。从市售的酵母多聚糖制剂中去除可能的活性物质的一种方法是用无热原水洗涤，以获得ZWIS(酵母多糖水不溶部分)、乙醇不溶(ETIS)或氯仿/甲醇不溶(CMIS)制剂。研究了不同洗涤的酵母多糖制剂对核因子-kappaB活化及与β-葡聚糖识别蛋白结合的影响。表达Toll样受体(TLR)2的HEK293细胞可显著激活核因子-kappaB，并通过TLR2和β-葡聚糖受体Dectin-1的共表达增强核因子-kappaB的活性。然而，酵母多糖制剂与Dectin-1蛋白的结合能力即使在TLR2介导的NF-kappa B活性被取消的有机溶剂处理后也没有改变。另一条涉及CARD9/Bc110的核因子-kappa B激活通路是在Dectin-1存在的情况下由这些酵母多糖制剂触发的。结果表明，酵母多糖的β-葡聚糖依赖特性不受氯仿/甲醇或乙醇洗涤的影响，TLR2介导的活性很容易被这些有机溶剂消除。在研究真菌大分子的天然免疫反应时，这种处理方法可能有助于区分TLR2和β-葡聚糖受体的天然配体。
Zymosan activates phagocytes through the innate immune system and causes inflammatory responses in animals. Because of the complexity of the active substances included in Zymosan preparations, simplifying the active moiety actually responsible for innate immune recognition is needed. One way to remove possible active substances from commercially available Zymosan preparations is to wash then with pyrogen-free water to obtain a ZWIS (Zymosan water insoluble fraction), ethanol insoluble (EtIS), or chloroform/methanol insoluble (CMIS) preparation. The effects of various washed Zymosan preparations on nuclear factor (NF)-kappa B activation and binding to beta-glucan recognition protein were examined. Significant NF-kappa B activation by Toll-like receptor (TLR) 2-expressing HEK293 cells and enhanced NF-kappa B activity via the co-expression of TLR2 and Dectin-1, a functional P-glucan receptor, was only observed in response to ZWIS. However, the ability of Zymosan preparations to bind Dectin-1 protein was not altered even after treatment with the organic solvents by which the TLR2-mediated NF-kappa B activity was abolished. Another NF-kappa B activation pathway involving CARD9/Bc110 was triggered by these Zymosan preparations in the presence of Dectin-1. The results suggest that the beta-glucan-dependent characteristics of Zymosan were not affected by the washing with chloroform/methanol or ethanol, and that TLR2-mediated activity was easily eliminated with these organic solvents. This treatment might be useful for distinguishing natural ligands for TLR2 and beta-glucan receptors when studying the innate immune response to fungal macromolecules.