中国大鲵（Andrias davidianus）中Intlectin-1的分子克隆、表达分析及功能鉴定
Molecular cloning, expression analysis, and functional characterization of Intelectin-1 from Chinese giant salamanders (Andrias davidianus).
作者信息
Mo Haolin, Liu Kexin, An Xiaoran, Chen Yongqing, Yu Jiajia, Yu Huixia, Yao Mingxing, Song Weijia, Li Yang, Wang Lixin
机构信息
College of Animal Science and Technology, Northwest A&F University, Xinong Road, Yangling, 712100, Shaanxi, China.
Fisheries and Fisheries Administration of Department of Agriculture and Rural Affairs of Shaanxi Province, Xi'an, 710003, Shaanxi, China.
出版信息
Genes Genomics. 2025 Mar 18. doi: 10.1007/s13258-025-01630-1.
BACKGROUND
Intelectin (ITLN) plays a pivotal role in innate immunity, inflammatory responses, and tumor progression. However, its physiological roles in caudate amphibians, such as Andrias davidianus, remain elusive.
OBJECTIVE
A proto-type of intelectin (AdITLN1) from A. davidianus was characterized, following which an AdITLN1 homology model was generated to analyze its expression profiles and functional characterizations.
METHODS
The intelectin1 gene (AdITLN1) was cloned, and its evolutionary relationship with ITLN1 from other species was explored. Additionally, a homology model was generated using Molecular Operating Environment (MOE) software, and the active binding pocket of AdITLN1 was identified. Infections with Aeromonas hydrophila were conducted to analyze changes in ITLN1 transcript levels in liver tissue. Finally, recombinant A. davidianus ITLN1 protein (rAdITLN1) was synthesized using prokaryotic expression, and its bacterial agglutination activity and impact on macrophage phagocytosis were examined.
RESULTS
Multiple sequence alignment and phylogenetic analysis indicated that AdITLN1 shared the closest evolutionary relationship with amphibians, with its structure being similar to that of human intelectin1 and Xenopus Embryonic Epidermal Lectin. Moreover, AdITLN1 mRNA was expressed in a wide range of tissues and was significantly up-regulated post-A. hydrophila infection. Meanwhile, the AdITLN1 protein was successfully expressed and purified in Escherichia coli (E. coli) BL21 (DE3). The recombinant AdITLN1 (rAdITLN1) displayed strong agglutination activity against different Gram-negative and Gram-positive bacteria. Lastly, The phagocytosis of rAdITLN1-treated E. coli by macrophages was significantly enhanced.
CONCLUSION
The results of the present study demonstrated that AdITLN1 was a multifunctional immune protein with potent immunomodulatory activity. This study offers valuable insights into disease control in giant salamanders and the conservation of natural resources.
背景
凝集素（ITLN）在天然免疫、炎症反应和肿瘤进展中起关键作用。然而，其在尾状两栖动物如大鲵中的生理作用仍不清楚。
目的
对大鲵的一种凝集素原型（AdITLN1）进行表征，随后构建AdITLN1同源模型以分析其表达谱和功能特性。
方法
克隆凝集素1基因（AdITLN1），并探索其与其他物种ITLN1的进化关系。此外，使用分子操作环境（MOE）软件构建同源模型，并鉴定AdITLN1的活性结合口袋。进行嗜水气单胞菌感染实验以分析肝组织中ITLN1转录水平的变化。最后，利用原核表达合成重组大鲵ITLN1蛋白（rAdITLN1），并检测其细菌凝集活性及对巨噬细胞吞噬作用的影响。
结果
多序列比对和系统发育分析表明，AdITLN1与两栖动物的进化关系最密切，其结构与人类凝集素1和非洲爪蟾胚胎表皮凝集素相似。此外，AdITLN1 mRNA在多种组织中表达，在嗜水气单胞菌感染后显著上调。同时，AdITLN1蛋白在大肠杆菌BL21（DE3）中成功表达并纯化。重组AdITLN1（rAdITLN1）对不同革兰氏阴性菌和革兰氏阳性菌均表现出较强的凝集活性。最后，rAdITLN1处理的大肠杆菌被巨噬细胞吞噬作用显著增强。
结论
本研究结果表明，AdITLN1是一种具有强大免疫调节活性的多功能免疫蛋白。该研究为大鲵疾病防控和自然资源保护提供了有价值的见解。
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