Key Laboratory of Marine Genetics and Breeding, Ministry of Education, College of Marine Life Sciences, Ocean University of China, Qingdao, Shandong, China.
Key Laboratory of Marine Genetics and Breeding, Ministry of Education, College of Marine Life Sciences, Ocean University of China, Qingdao, Shandong, China; Department of Life Science and Engineering, Jining University, Jining, China.
Fish Shellfish Immunol. 2019 Sep;92:11-20. doi: 10.1016/j.fsi.2019.05.054. Epub 2019 May 24.
C-type lectin is a type of carbohydrate-binding protein and plays significant roles in innate immune response against pathogen infection. To date, thousands of C-type lectin had been identified in teleost. In the present study, we isolated a novel isoform of C-type lectin (OppCTL) from spotted knifejaw (Oplegnathus punctatus). The OppCTL encoded a typical Ca-dependent carbohydrate-binding protein, and was mainly expressed in liver in a tissue specific fashion. The expression of OppCTL was significantly up-regulated following Vibrio anguillarum infection in vivo, suggesting involvement in immune response. Hemagglutination analysis showed that the recombinant OppCTL (rOppCTL) could agglutinate erythrocyte from Mus musculus, Oplegnathus punctatus, Sebastes schlegelii and Paralichthys olivaceus. The rOppCTL could bind and agglutinate all tested bacteria. The rOppCTL possessed capacities of calcium-dependent agglutination to all tested bacteria. Sugar binding assay revealed that rOppCTL could also bind to the glycoconjugates of the bacterial surface, including lipopolysaccharide and peptidoglycan. Interestingly, Dual-luciferase analysis revealed that OppCTL could inhibit the activity of NF-κB in HEK-293T cells after OppCTL overexpression. Taken together, these results indicate that OppCTL has immune activity capable of defending invading pathogens and possesses potential immunoregulatory activity, enriching our understanding of the function of C-type lectin.
C 型凝集素是一种糖结合蛋白,在先天免疫反应中对病原体感染起着重要作用。迄今为止,已在硬骨鱼类中鉴定出数千种 C 型凝集素。本研究从斑点叉尾鮰(Oplegnathus punctatus)中分离出一种新型 C 型凝集素(OppCTL)。OppCTL 编码一种典型的 Ca 依赖性糖结合蛋白,在组织中特异性地主要在肝脏中表达。OppCTL 的表达在体内感染鳗弧菌后显著上调,提示其参与免疫反应。血凝分析表明,重组 OppCTL(rOppCTL)可凝集来自 Mus musculus、Oplegnathus punctatus、Sebastes schlegelii 和 Paralichthys olivaceus 的红细胞。rOppCTL 可结合并凝集所有测试的细菌。rOppCTL 具有对所有测试细菌的钙离子依赖性凝集能力。糖结合分析表明,rOppCTL 还可以与细菌表面的糖缀合物结合,包括脂多糖和肽聚糖。有趣的是,双荧光素酶分析显示,OppCTL 过表达后可抑制 HEK-293T 细胞中 NF-κB 的活性。综上所述,这些结果表明 OppCTL 具有抵御入侵病原体的免疫活性,并具有潜在的免疫调节活性,丰富了我们对 C 型凝集素功能的理解。