Yao Tuo, Tong Zhengxin, Lu Jie, Fu Shengli, Cheng Changhong, Ye Lingtong
Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, China.
Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, China; College of Marine Science and Environment, Dalian Ocean University, Dalian, 116023, China.
Fish Shellfish Immunol. 2024 Dec;155:110029. doi: 10.1016/j.fsi.2024.110029. Epub 2024 Nov 17.
C-type lectins (CTLs), a member of pattern recognition receptors, play an important role in the innate immunity by recognizing invading microorganisms. In this study, a novel perlucin gene (designated as HdPer 3), a typical CTLs was cloned and characterized from the small abalone Haliotis diversicolor. The open reading frame of HdPer 3 was 471 bp, encoding a protein of 156 amino acids that included a single carbohydrate-recognition domain. HdPer 3 was widely expressed in all tested tissues and developmental stage. HdPer 3 expression was significantly up-regulated after Vibrio harveyi infection, suggesting that HdPer 3 was activated in response to bacterial infection. The encapsulation ability of hemocytes could be significantly enhanced by the recombinant protein HdPer 3 (rHdPer 3). To understand the regulation mechanism of the HdPer 3, HdPer 3 was silenced in vivo by RNAi. Knocking down HdPer 3 decreased the hemocytes phagocytosis. Meanwhile, knocking down HdPer 3 can reduce the expression of 2 phagocytosis-related genes (Rab and Dynamin), TNF-α, and 2 MAPK pathway-related genes (MAPK-X1 and Ras) after V. harveyi infection. Moreover, HdPer 3 interference could increase the bacterial load in the hemolymph and the mortality of abalones after V. harveyi infection. All these results suggested that HdPer 3 played a crucial role in the defense against V. harveyi infection by recognizing bacterial pathogens and activating the expression of immune-related genes.
C型凝集素(CTLs)是模式识别受体的成员之一,通过识别入侵的微生物在先天免疫中发挥重要作用。在本研究中,从杂色鲍中克隆并鉴定了一个新的穿孔蛋白基因(命名为HdPer 3),它是一种典型的C型凝集素。HdPer 3的开放阅读框为471 bp,编码一个156个氨基酸的蛋白质,该蛋白质包含一个单一的碳水化合物识别结构域。HdPer 3在所有测试组织和发育阶段均广泛表达。哈维氏弧菌感染后,HdPer 3的表达显著上调,表明HdPer 3在细菌感染后被激活。重组蛋白HdPer 3(rHdPer 3)可显著增强血细胞的包囊化能力。为了解HdPer 3的调控机制,通过RNA干扰在体内使HdPer 3沉默。敲低HdPer 3会降低血细胞的吞噬作用。同时,敲低HdPer 3可降低哈维氏弧菌感染后2个吞噬相关基因(Rab和发动蛋白)、TNF-α以及2个丝裂原活化蛋白激酶(MAPK)途径相关基因(MAPK-X1和Ras)的表达。此外,HdPer 3干扰会增加哈维氏弧菌感染后血淋巴中的细菌载量以及鲍鱼的死亡率。所有这些结果表明,HdPer 3通过识别细菌病原体并激活免疫相关基因的表达,在抵御哈维氏弧菌感染中发挥关键作用。