The JNK/NFκB pathway is required to activate murine lymphocytes induced by a sulfated polysaccharide from Ecklonia cava
The JNK/NFκB pathway is required to activate murine lymphocytes induced by a sulfated polysaccharide from Ecklonia cava
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DOI：
10.1016/j.bbagen.2012.12.008
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发表时间：
2013-03-01
影响因子：
3
通讯作者：
Jee, Youngheun
中科院分区：
文献类型：
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作者：
Ahn, Ginnae;Bing, So Jin;Jee, Youngheun
背景：静脉昆布 (E. cava) 已被证实的免疫调节和免疫系统激活特性归因于其丰富的多糖含量。因此，我们研究了静脉曲张的硫酸多糖（SP）是否特异性激活蛋白激酶（MAPK）和核因子κB（NF kappa B）来激发免疫反应。方法：为了评估免疫反应，从ICR小鼠的脾脏中分离淋巴细胞，并与SP及其抑制剂一起培养。检测方法包括 H-3-胸苷掺入、流式细胞术、实时聚合酶链式反应 (rtPCR)、酶联免疫吸附测定 (ELISA)、细胞内细胞因子测定、蛋白质印迹和电泳迁移率变动测定 (EMSA)。 结果：SP 剂量依赖性地增加淋巴细胞的增殖，但没有细胞毒性。特别是，SP显着增强CD3(+)成熟T细胞和CD45R/B220(+)泛B细胞的增殖和分化。此外。与未处理的细胞相比，SP 增加了 IL-2、IgG(1a) 和 IgG(2b) 的表达和/或产生。随后应用 JNK (SP600125)、NF kappa B (PDTC) 和丝氨酸蛋白酶 (TPCK) 抑制剂显着抑制 SP 处理的淋巴细胞的增殖和 IL-2 产生，以及 JNK 和 I kappa B 的磷酸化、核 NF kappa B p65 的激活以及 NF kappa B p65 DNA 的结合。此外，即使在 SP 存在的情况下，JNK 和 NF kappa B 抑制剂的共同应用也能完全阻断淋巴细胞的增殖。结论：这些结果表明 SP 通过 JNK 和 NF kappa B 途径诱导 T 和 B 细胞反应。一般意义：这里首次检测了 SP 对脾淋巴细胞活化的影响，并表明了潜在的功能机制。皇冠版权所有 (C) 2013 由 Elsevier B.V. 出版。保留所有权利。
Background: The proven immunomodulatory and immune system activating properties of Ecklonia cava (E. cava) have been attributed to its plentiful polysaccharide content. Therefore, we investigated whether the sulfated polysaccharide (SP) of E. cava specifically activates the protein kinases (MAPKs) and nuclear factor-kappa B (NF kappa B) to incite immune responses.Methods: To assess immune responsiveness, lymphocytes were isolated from spleens of ICR mice and cultured with SP and its inhibitors. Assays included H-3-thymidine incorporation, flow cytometry, real time polymerase chain reaction (rtPCR), enzyme linked immunosorbent assay (ELISA), intracellular cytokine assay, Western blot, and electrophoretic mobility shift assay (EMSA).Results: SP dose-dependently increased the proliferation of lymphocytes without cytotoxicity. In particular, SP markedly enhanced the proliferation and differentiation of CD3(+) mature T cells and CD45R/B220(+) pan B cells. Additionally. SP increased the expression and/or production of IL-2, IgG(1a), and IgG(2b) compared to that in untreated cells. The subsequent application of JNK (SP600125), NF kappa B (PDTC), and serine protease (TPCK) inhibitors significantly inhibited the proliferation and IL-2 production of SP-treated lymphocytes as well as the phosphorylation of JNK and I kappa B, the activation of nuclear NF kappa B p65, and binding of NF kappa B p65 DNA. Moreover, co-application of both JNK and NF kappa B inhibitors completely blocked the proliferation of lymphocytes even in the presence of SP.Conclusion: These results suggest that SP induced T and B cell responses via both JNK and NF kappa B pathways.General significance: The effect of SP on splenic lymphocyte activation was assayed here for the first time and indicated the underlying functional mechanism. Crown Copyright (C) 2013 Published by Elsevier B.V. All rights reserved.