An invertebrate β-integrin mediates coelomocyte phagocytosis via activation of septin2 and 7 but not septin10.
An invertebrate β-integrin mediates coelomocyte phagocytosis via activation of septin2 and 7 but not septin10.
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DOI：
10.1016/j.ijbiomac.2018.03.033
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发表时间：
2018-07
影响因子：
8.2
通讯作者：
Zhenhui Wang;Zhimeng Lv;Chenghua Li;Y. Shao;Weiwei Zhang;Xuelin Zhao
中科院分区：
文献类型：
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作者：
Zhenhui Wang;Zhimeng Lv;Chenghua Li;Y. Shao;Weiwei Zhang;Xuelin Zhao
我们以前已经证实，β-整合素从仿刺参（命名为AjITGB）结合LPS和介导的免疫反应。在本研究中，我们发现AjITGB正性地促进位于体腔内的棘皮动物免疫细胞（指定为体腔细胞）的吞噬活性。流式细胞仪检测显示，AjITGB siRNA体外和体内干扰后，吞噬细胞百分率分别显著降低37.3%和41.36%，这一结果与抗AjITGB抗体阻断治疗后观察到的降低一致。这些降低的吞噬活性通过rAjITGB补充部分恢复。为了更好地理解这种吞噬作用的分子机制，三个吞噬相关蛋白，包括Ajseptin 2，Ajseptin 7和Ajseptin 10，被克隆并进一步表征。AjITGB和Ajseptin 2/7在mRNA和蛋白质水平上检测到完全一致的表达谱，inV。海参此外，Ajseptin 2/7表达水平在AjITGB沉默组中显著下调，并且在rAjITGB施用后可以部分恢复到其原始水平。然而，Ajseptin 10在相同条件下未显示显著变化。吞噬试验进一步表明，Ajseptin 2/7，而不是Ajseptin 10，是至关重要的介导体腔细胞吞噬。通过特异性siRNA敲低三个基因表明Ajseptin 2/7显著降低体腔细胞吞噬作用，并且这种降低在rAjITGB补充后完全恢复。在AjSEPT 10 siRNA干扰组和rAjITGB补充组中，吞噬率没有显著变化。我们的所有结果证实AjITGB调节Ajseptin 2/7而不是AjSEPT 10的体腔细胞吞噬活性，并进一步支持Ajseptins在AjITGB介导的吞噬作用中的不同作用。
We have previously confirmed that β-integrin fromApostichopus japonicus(designated AjITGB) binds LPS and mediates the immune response. In this study, we found that AjITGB positively promoted the echinoderm immune cells located in the coelomic cavity (designated as coelomocyte) phagocytic activities. Flow cytometry assay indicated that the phagocytic percentage was significantly decreased, by a 37.3% and 41.36%, after AjITGB siRNA inferencein vitroandin vivo, respectively, a result consistent with the decrease observed with an anti-AjITGB antibody blocking treatment. These decreased phagocytic activities were partially recovered by rAjITGB supplementation. To better understand the molecular mechanism underlying this phagocytosis, three phagocytic-related proteins, including Ajseptin2, Ajseptin7 and Ajseptin10, were cloned and further characterized. Completely consistent expression profiles were detected between AjITGB and Ajseptin2/7, at both the mRNA and protein levels, inV. splendidus-challenged sea cucumber. Furthermore, Ajseptin2/7 expression levels were significantly down-regulated in the AjITGB silencing group and could be partially recovered to its original level after rAjITGB administration. However, Ajseptin10 displayed no significant change in the same condition. A phagocytic assay further indicated that Ajseptin2/7, but not Ajseptin10, was crucial in mediating coelomocyte phagocytosis. The knockdown of the three genes by specific siRNAs indicated that Ajseptin2/7 significantly decreased coelomocyte phagocytosis, and this decrease was completely recovered after rAjITGB supplementation. There was no significant change in the phagocytosis rate in both the AjSEPT10 siRNA interference and rAjITGB supplementation groups. All our results confirmed that AjITGB modulates coelomocyte phagocytosisviathe activation of Ajseptin2/7 but not AjSEPT10 and further supported the divergent roles of Ajseptins in AjITGB-mediated phagocytosis.