PaCLR介导PaLECT2激活脂多糖刺激的香鱼(Plecoglossus altivelis)单核巨噬细胞功能
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摘要
香鱼(Plecoglossus altivelis)白细胞衍生趋化因子2(leukocyte cell-derived chemotaxin 2,PaLECT2)可以激活香鱼头肾来源的单核巨噬细胞(monocytes/macrophages,MO/MΦ),提高细菌感染后香鱼存活率。进一步研究显示,PaLECT2与香鱼C型凝集素受体(C-type lectin receptor,PaCLR)相互作用可激活静息状态下香鱼MO/MΦ,增强其免疫活性。在前期基础上,本研究通过抗体封闭实验检测PaCLR是否介导PaLECT2激活LPS刺激的MO/MΦ功能。结果显示,重组PaLECT2成熟肽(rPaLECT2m)显著提高了LPS刺激的香鱼MO/MΦ细胞因子的表达,并增强了其吞噬能力和杀菌活性。封闭PaCLR显著抑制了rPaLECT2m激活LPS刺激的香鱼MO/MΦ细胞因子表达、吞噬和杀菌功能。因此,PaCLR介导PaLECT2可以激活病理状态香鱼MO/MΦ。这一认识进一步揭示了LECT2在病理状态的鱼类巨噬细胞调控和炎症反应中的作用及机制。
Leukocyte cell-derived chemotaxin 2 of ayu Plecoglossus altivelis(Pa LECT2) is shown to activate ayu head kidney-derived monocytes/macrophages(MO/MΦ) in vitro to improve the cure of bacterial-infected fish. Our earlier research reveals that Pa LECT2 could activate the resting MO/MΦ and enhance its immuno-activity by interacting with a C-type lectin receptor of ayu(Pa CLR), based on which we examined the effect of Pa CLR on Pa LECT2 activation of LPS-stimulated ayu MO/MΦ using antibody neutralization method. Results show that the recombinant Pa LECT2 mature peptide(r Pa LECT2m) increased the cytokine expression, and enhanced phagocytosis and bactericidal activity of LPS-stimulated ayu MO/MΦ. By blocking the ectodomain of Pa CLR by anti-Pa CLR, these effects were inhibited significantly. Therefore, Pa CLR could mediate the activation effect of Pa LECT2 on LPS stimulated MO/MΦ. This finding revealed the mechanism of LECT2 involvement in the regulation and inflammatory response of the fish monocyte/macrophage.
Leukocyte cell-derived chemotaxin 2 of ayu Plecoglossus altivelis(Pa LECT2) is shown to activate ayu head kidney-derived monocytes/macrophages(MO/MΦ) in vitro to improve the cure of bacterial-infected fish. Our earlier research reveals that Pa LECT2 could activate the resting MO/MΦ and enhance its immuno-activity by interacting with a C-type lectin receptor of ayu(Pa CLR), based on which we examined the effect of Pa CLR on Pa LECT2 activation of LPS-stimulated ayu MO/MΦ using antibody neutralization method. Results show that the recombinant Pa LECT2 mature peptide(r Pa LECT2m) increased the cytokine expression, and enhanced phagocytosis and bactericidal activity of LPS-stimulated ayu MO/MΦ. By blocking the ectodomain of Pa CLR by anti-Pa CLR, these effects were inhibited significantly. Therefore, Pa CLR could mediate the activation effect of Pa LECT2 on LPS stimulated MO/MΦ. This finding revealed the mechanism of LECT2 involvement in the regulation and inflammatory response of the fish monocyte/macrophage.
引文
施晓峰,曲朦,王航俊等,2010.赤点石斑鱼LECT2基因的克隆与组织表达分析.厦门大学学报(自然科学版),49(5):694-700
Ao J Q,Ding Y,Chen Y Y et al,2015.Molecular characterization and biological effects of a C-type lectin-like receptor in large yellow croaker(Larimichthys crocea).International Journal of Molecular Sciences,16(12):29631-29642
Chen C K,Yang C Y,Hua K T et al,2014a.Leukocyte cell-derived chemotaxin 2 antagonizes MET receptor activation to suppress hepatocellular carcinoma vascular invasion by protein tyrosine phosphatase 1B recruitment.Hepatology,59(3):974-985
Chen J,Chen Q,Lu X J et al,2014b.LECT2 improves the outcomes in ayu with Vibrio anguillarum infection via monocytes/macrophages.Fish&Shellfish Immunology,41(2):586-592
Chen J,Lu X J,Yang H Y et al,2010.An interaction between a C-type lectin receptor and leukocyte cell-derived chemotaxin 2 of ayu,Plecoglossus altivelis.Fish&Shellfish Immunology,28(1):245-248
Chen S X,Ma H L,Shi Y H et al,2016.Molecular and functional characterization of a novel CD302 gene from ayu(Plecoglossus altivelis).Fish&Shellfish Immunology,55:140-148
Fujiki K,Shin D H,Nakao M et al,2000.Molecular cloning of carp(Cyprinus carpio)leucocyte cell-derived chemotaxin 2,glia maturation factorβ,CD45 and lysozyme C by use of suppression subtractive hybridisation.Fish&Shellfish Immunology,10(7):643-650
Kerrigan A M,Brown G D,2009.C-type lectins and phagocytosis.Immunobiology,214(7):562-575
Kokkinos P A,Kazantzi A,Sfyroera G et al,2005.Molecular cloning of leukocyte cell-derived chemotaxin 2 in rainbow trout.Fish&Shellfish Immunology,18(5):371-380
Lan F,Misu H,Chikamoto K et al,2014.LECT2 functions as a hepatokine that links obesity to skeletal muscle insulin resistance.Diabetes,63(5):1649-1664
Li M Y,Chen J,Shi Y H,2008.Molecular cloning of leucocyte cell-derived chemotaxin-2 gene in croceine croaker(Pseudosciaena crocea).Fish&Shellfish Immunology,24(2):252-256
Lin A F,Xiang L X,Wang Q L et al,2009.The dc-sign of zebrafish:insights into the existence of a CD209 homologue in a lower vertebrate and its involvement in adaptive immunity.Journal of Immunology,183(11):7398-7410
Lin B,Chen S W,Cao Z et al,2007.Acute phase response in zebrafish upon Aeromonas salmonicida and Staphylococcus aureus infection:striking similarities and obvious differences with mammals.Molecular Immunology,44(4):295-301
Lu X J,Chen J,Yu C H et al,2013a.LECT2 protects mice against bacterial sepsis by activating macrophages via the CD209a receptor.The Journal of Experimental Medicine,210(1):5-13
Lu X J,Chen Q,Rong Y J et al,2016.LECT2 drives haematopoietic stem cell expansion and mobilization via regulating the macrophages and osteolineage cells.Nature Communications,7:12719
Lu X J,Hang X Y,Yin L et al,2013b.Sequencing of the first ayu(Plecoglossus altivelis)macrophage transcriptome and microarray development for investigation the effect of LECT2 on macrophages.Fish&Shellfish Immunology,34(2):497-504
Ma H L,Shi Y H,Zhang X H et al,2016.A transmembrane C-type lectin receptor mediates LECT2 effects on head kidney-derived monocytes/macrophages in a teleost,Plecoglossus altivelis.Fish&Shellfish Immunology,51:70-76
Morris MC,Gilliam EA,Li L,2015.Innate immune programing by endotoxin and its pathological consequences.Frontiers in Immunology,5:680
Saito T,Okumura A,Watanabe H et al,2004.Increase in hepatic NKT cells in leukocyte cell-derived chemotaxin 2-deficient mice contributes to severe concanavalin a-induced hepatitis.Journal of Immunology,173(1):579-585
Soanes K H,Figuereido K,Richards R C et al,2004.Sequence and expression of C-type lectin receptors in atlantic salmon(Salmo salar).Immunogenetics,56(8):572-584
Swain P,Nayak S K,Nanda P K et al,2008.Biological effects of bacterial lipopolysaccharide(endotoxin)in fish:a review.Fish&Shellfish Immunology,25(3):191-201
Wei J G,Guo M L,Cui H C et al,2011.A new leukocyte cell-derived chemotaxin-2 from marine fish grouper,Epinephelus coioides:molecular cloning and expression analysis.Fish&Shellfish Immunology,31(4):600-605
Yamagoe S,Yamakawa Y,Matsuo Y et al,1996.Purification and primary amino acid sequence of a novel neutrophil chemotactic factor LECT2.Immunology Letters,52(1):9-13
Yang G J,Lu X J,Chen Q et al,2015.Molecular characterization and functional analysis of a novel C-type lectin receptor-like gene from a teleost fish,Plecoglossus altivelis.Fish&Shellfish Immunology,44(2):603-610
Zhang R C,Chen J,Li C H et al,2011.Prokaryotic expression,purification,and refolding of leukocyte cell-derived chemotaxin 2 and its effect on gene expression of head kidney-derived macrophages of a teleost fish,ayu(Plecoglossus altivelis).Fish&Shellfish Immunology,31(6):911-918
Zhang X H,Shi Y H,Chen J,2015.Molecular characterization of a transmembrane C-type lectin receptor gene from ayu(Plecoglossus altivelis)and its effect on the recognition of different bacteria by monocytes/macrophages.Molecular Immunology,66(2):439-450
Ao J Q,Ding Y,Chen Y Y et al,2015.Molecular characterization and biological effects of a C-type lectin-like receptor in large yellow croaker(Larimichthys crocea).International Journal of Molecular Sciences,16(12):29631-29642
Chen C K,Yang C Y,Hua K T et al,2014a.Leukocyte cell-derived chemotaxin 2 antagonizes MET receptor activation to suppress hepatocellular carcinoma vascular invasion by protein tyrosine phosphatase 1B recruitment.Hepatology,59(3):974-985
Chen J,Chen Q,Lu X J et al,2014b.LECT2 improves the outcomes in ayu with Vibrio anguillarum infection via monocytes/macrophages.Fish&Shellfish Immunology,41(2):586-592
Chen J,Lu X J,Yang H Y et al,2010.An interaction between a C-type lectin receptor and leukocyte cell-derived chemotaxin 2 of ayu,Plecoglossus altivelis.Fish&Shellfish Immunology,28(1):245-248
Chen S X,Ma H L,Shi Y H et al,2016.Molecular and functional characterization of a novel CD302 gene from ayu(Plecoglossus altivelis).Fish&Shellfish Immunology,55:140-148
Fujiki K,Shin D H,Nakao M et al,2000.Molecular cloning of carp(Cyprinus carpio)leucocyte cell-derived chemotaxin 2,glia maturation factorβ,CD45 and lysozyme C by use of suppression subtractive hybridisation.Fish&Shellfish Immunology,10(7):643-650
Kerrigan A M,Brown G D,2009.C-type lectins and phagocytosis.Immunobiology,214(7):562-575
Kokkinos P A,Kazantzi A,Sfyroera G et al,2005.Molecular cloning of leukocyte cell-derived chemotaxin 2 in rainbow trout.Fish&Shellfish Immunology,18(5):371-380
Lan F,Misu H,Chikamoto K et al,2014.LECT2 functions as a hepatokine that links obesity to skeletal muscle insulin resistance.Diabetes,63(5):1649-1664
Li M Y,Chen J,Shi Y H,2008.Molecular cloning of leucocyte cell-derived chemotaxin-2 gene in croceine croaker(Pseudosciaena crocea).Fish&Shellfish Immunology,24(2):252-256
Lin A F,Xiang L X,Wang Q L et al,2009.The dc-sign of zebrafish:insights into the existence of a CD209 homologue in a lower vertebrate and its involvement in adaptive immunity.Journal of Immunology,183(11):7398-7410
Lin B,Chen S W,Cao Z et al,2007.Acute phase response in zebrafish upon Aeromonas salmonicida and Staphylococcus aureus infection:striking similarities and obvious differences with mammals.Molecular Immunology,44(4):295-301
Lu X J,Chen J,Yu C H et al,2013a.LECT2 protects mice against bacterial sepsis by activating macrophages via the CD209a receptor.The Journal of Experimental Medicine,210(1):5-13
Lu X J,Chen Q,Rong Y J et al,2016.LECT2 drives haematopoietic stem cell expansion and mobilization via regulating the macrophages and osteolineage cells.Nature Communications,7:12719
Lu X J,Hang X Y,Yin L et al,2013b.Sequencing of the first ayu(Plecoglossus altivelis)macrophage transcriptome and microarray development for investigation the effect of LECT2 on macrophages.Fish&Shellfish Immunology,34(2):497-504
Ma H L,Shi Y H,Zhang X H et al,2016.A transmembrane C-type lectin receptor mediates LECT2 effects on head kidney-derived monocytes/macrophages in a teleost,Plecoglossus altivelis.Fish&Shellfish Immunology,51:70-76
Morris MC,Gilliam EA,Li L,2015.Innate immune programing by endotoxin and its pathological consequences.Frontiers in Immunology,5:680
Saito T,Okumura A,Watanabe H et al,2004.Increase in hepatic NKT cells in leukocyte cell-derived chemotaxin 2-deficient mice contributes to severe concanavalin a-induced hepatitis.Journal of Immunology,173(1):579-585
Soanes K H,Figuereido K,Richards R C et al,2004.Sequence and expression of C-type lectin receptors in atlantic salmon(Salmo salar).Immunogenetics,56(8):572-584
Swain P,Nayak S K,Nanda P K et al,2008.Biological effects of bacterial lipopolysaccharide(endotoxin)in fish:a review.Fish&Shellfish Immunology,25(3):191-201
Wei J G,Guo M L,Cui H C et al,2011.A new leukocyte cell-derived chemotaxin-2 from marine fish grouper,Epinephelus coioides:molecular cloning and expression analysis.Fish&Shellfish Immunology,31(4):600-605
Yamagoe S,Yamakawa Y,Matsuo Y et al,1996.Purification and primary amino acid sequence of a novel neutrophil chemotactic factor LECT2.Immunology Letters,52(1):9-13
Yang G J,Lu X J,Chen Q et al,2015.Molecular characterization and functional analysis of a novel C-type lectin receptor-like gene from a teleost fish,Plecoglossus altivelis.Fish&Shellfish Immunology,44(2):603-610
Zhang R C,Chen J,Li C H et al,2011.Prokaryotic expression,purification,and refolding of leukocyte cell-derived chemotaxin 2 and its effect on gene expression of head kidney-derived macrophages of a teleost fish,ayu(Plecoglossus altivelis).Fish&Shellfish Immunology,31(6):911-918
Zhang X H,Shi Y H,Chen J,2015.Molecular characterization of a transmembrane C-type lectin receptor gene from ayu(Plecoglossus altivelis)and its effect on the recognition of different bacteria by monocytes/macrophages.Molecular Immunology,66(2):439-450