BLP耐受通过上调MARCO增强巨噬细胞的吞噬能力
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摘要
目的:探讨胶原样结构巨噬细胞受体(MARCO)在细菌脂蛋白(BLP)耐受巨噬细胞吞噬能力增强过程中的作用。方法:分离、分化和培养骨髓来源的巨噬细胞(BMDMs),并制备BLP耐受细胞;利用流式细胞术和荧光显微镜检测并比较BLP耐受与非耐受细胞吞噬细菌的能力,同时用流式细胞术和qPCR检测BLP耐受对MARCO蛋白及mRNA表达的影响;进一步利用小干扰RNA下调MARCO表达,通过流式细胞术及荧光技术探讨MARCO对BLP耐受巨噬细胞吞噬能力的影响。结果:BLP耐受巨噬细胞吞噬细菌的量较非耐受巨噬细胞明显增加(P<0.05);同时BLP耐受巨噬细胞膜表面MARCO蛋白表达水平较非耐受细胞明显升高,并且在细菌刺激后进一步增加,MARCO的mRNA水平变化与蛋白水平变化一致;下调MARCO表达后,BLP耐受巨噬细胞对细菌的吞噬能力明显下降,提示BLP耐受巨噬细胞吞噬能力增强与巨噬细胞膜表面MARCO蛋白的表达上调有关。结论:BLP耐受通过上调巨噬细胞膜表面MARCO蛋白的表达,增强其对细菌的吞噬能力。
AIM: To investigate the role of macrophage receptor with collagenous structure(MARCO), class A scavenger receptor, in enhancing the phagocytosis of bacterial lipoprotein(BLP) tolerized macrophages. METHODS: Bone marrow derived macrophages(BMDMs) was differentiated and cultured. BLP-tolerized macrophages were induced by treating the BMDMs with BLP for 24 h while the naive BMDMs were unstimulated. The phagocytic capacities of BLP-tolerized BMDMs and the naive BMDMs were detected by flow cytometry and fluorescence microscopy. Meanwhile, the effect of BLP tolerance on MARCO protein and mRNA expression was assessed by flow cytometry and qPCR. Furthermore, the small interference RNA was applied to down-regulate the expression of MARCO, and the effect of MARCO on the phagocytosis was measured by flow cytometry and fluorescent technique. RESULTS: Compared with the naive BMDMs, a prominently larger quantity of bacteria was engulfed by BLP-tolerized BMDMs(P<0.05). In addition, the protein expression of MARCO significantly increased in BLP-tolerized BMDMs, and then the MARCO protein further increased after stimulated with bacteria. The mRNA level of MARCO was consistent with the protein. Moreover, after down-regulation of MARCO, the phagocytic capacity of BLP-tolerized BMDMs was noticed to decline conspicuously, indicating that the enhanced phagocytosis of BLP-tolerized BMDMs displayed a substantial correlation with the up-regulation of the MARCO expression on BLP-tolerized macrophages. CONCLUSION: BLP tolerance enhances the phagocytosis of BMDMs through the up-regulation of MARCO.
AIM: To investigate the role of macrophage receptor with collagenous structure(MARCO), class A scavenger receptor, in enhancing the phagocytosis of bacterial lipoprotein(BLP) tolerized macrophages. METHODS: Bone marrow derived macrophages(BMDMs) was differentiated and cultured. BLP-tolerized macrophages were induced by treating the BMDMs with BLP for 24 h while the naive BMDMs were unstimulated. The phagocytic capacities of BLP-tolerized BMDMs and the naive BMDMs were detected by flow cytometry and fluorescence microscopy. Meanwhile, the effect of BLP tolerance on MARCO protein and mRNA expression was assessed by flow cytometry and qPCR. Furthermore, the small interference RNA was applied to down-regulate the expression of MARCO, and the effect of MARCO on the phagocytosis was measured by flow cytometry and fluorescent technique. RESULTS: Compared with the naive BMDMs, a prominently larger quantity of bacteria was engulfed by BLP-tolerized BMDMs(P<0.05). In addition, the protein expression of MARCO significantly increased in BLP-tolerized BMDMs, and then the MARCO protein further increased after stimulated with bacteria. The mRNA level of MARCO was consistent with the protein. Moreover, after down-regulation of MARCO, the phagocytic capacity of BLP-tolerized BMDMs was noticed to decline conspicuously, indicating that the enhanced phagocytosis of BLP-tolerized BMDMs displayed a substantial correlation with the up-regulation of the MARCO expression on BLP-tolerized macrophages. CONCLUSION: BLP tolerance enhances the phagocytosis of BMDMs through the up-regulation of MARCO.
引文
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[9] Gursel M,Gursel I,Mostowski HS,et al.CXCL16 influences the nature and specificity of CpG-induced immune activation[J].J Immunol,2006,177(3):1575-1580.
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[12] van der Laan LJ,Dopp EA,Haworth R,et al.Regulation and functional involvement of macrophage scavenger receptor MARCO in clearance of bacteria in vivo[J].J Immunol,1999,162(2):939-947.
[13] Chen Y,Wermeling F,Sundqvist J,et al.A regulatory role for macrophage class A scavenger receptors in TLR4-mediated LPS responses[J].Eur J Immunol,2010,40(5):1451-1460.
[14] Matsushita N,Komine H,Grolleau-Julius A,et al.Targeting MARCO can lead to enhanced dendritic cell motility and anti-melanoma activity[J].Cancer Immunol Immunother,2010,59(6):875-884.
[15] Jing J,Yang IV,Hui L,et al.Role of macrophage receptor with collagenous structure in innate immune tolerance[J].J Immunol,2013,190(12):6360-6367.
[16] Schrum JE,Crabtree JN,Dobbs KR,et al.Cutting edge:plasmodium falciparum induces trained innate immunity[J].J Immunol,2018,200(4):1243-1248.
[17] Arredouani M,Yang Z,Ning Y,et al.The scavenger receptor MARCO is required for lung defense against pneumococcal pneumonia and inhaled particles[J].J Exp Med,2004,200(2):267-272.
[18] Benard EL,Roobol SJ,Spaink HP,et al.Phagocytosis of mycobacteria by zebrafish macrophages is dependent on the scavenger receptor Marco,a key control factor of pro-inflammatory signalling[J].Dev Comp Immunol,2014,47(2):223-233.
[19] Li Z,Jiao Y,Fan EK,et al.Aging-impaired filamentous actin polymerization signaling reduces alveolar macrophage phagocytosis of bacteria[J].J Immunol,2017,199(9):3176-3186.
[2] Liu J,Xiang J,Li X,et al.NF-κB activation is critical for bacterial lipoprotein tolerance-enhanced bactericidal activity in macrophages during microbial infection[J].Sci Rep,2017,7(1):40418.
[3] Bolland JVRA.IGG Fc receptors[J].Annu Rev Immunol,2001,19(1):275-290.
[4] van Lookeren Campagne M,Wiesmann C,Brown EJ.Macrophage complement receptors and pathogen clearance[J].Cell Microbiol,2007,9(9):2095-2102.
[5] 赵舒祺,蔡军伟,李雪,等.Rab7蛋白参与了BLP耐受巨噬细胞对细菌清除能力增强的过程[J].中国病理生理杂志,2016,32(5):900-906.
[6] Areschoug T,Gordon S.Scavenger receptors:role in innate immunity and microbial pathogenesis[J].Cell Microbiol,2009,11(8):1160-1169.
[7] Arredouani MS,Palecanda A,Koziel H,et al.MARCO is the major binding receptor for unopsonized particles and bacteria on human alveolar macrophages[J].J Immunol,2005,175(9):6058-6064.
[8] Mukhopadhyay S,Chen Y,Sankala M,et al.MARCO,an innate activation marker of macrophages,is a class A scavenger receptor for Neisseria meningitidis[J].Eur J Immunol,2006,36(4):940-949.
[9] Gursel M,Gursel I,Mostowski HS,et al.CXCL16 influences the nature and specificity of CpG-induced immune activation[J].J Immunol,2006,177(3):1575-1580.
[10] Doyle SE,O’Connell RM,Miranda GA,et al.Toll-like receptors induce a phagocytic gene program through p38[J].J Exp Med,2004,199(1):81-90.
[11] Mukhopadhyay S,Gordon S.The role of scavenger receptors in pathogen recognition and innate immunity[J].Immunobiology,2004,209(1-2):39-49.
[12] van der Laan LJ,Dopp EA,Haworth R,et al.Regulation and functional involvement of macrophage scavenger receptor MARCO in clearance of bacteria in vivo[J].J Immunol,1999,162(2):939-947.
[13] Chen Y,Wermeling F,Sundqvist J,et al.A regulatory role for macrophage class A scavenger receptors in TLR4-mediated LPS responses[J].Eur J Immunol,2010,40(5):1451-1460.
[14] Matsushita N,Komine H,Grolleau-Julius A,et al.Targeting MARCO can lead to enhanced dendritic cell motility and anti-melanoma activity[J].Cancer Immunol Immunother,2010,59(6):875-884.
[15] Jing J,Yang IV,Hui L,et al.Role of macrophage receptor with collagenous structure in innate immune tolerance[J].J Immunol,2013,190(12):6360-6367.
[16] Schrum JE,Crabtree JN,Dobbs KR,et al.Cutting edge:plasmodium falciparum induces trained innate immunity[J].J Immunol,2018,200(4):1243-1248.
[17] Arredouani M,Yang Z,Ning Y,et al.The scavenger receptor MARCO is required for lung defense against pneumococcal pneumonia and inhaled particles[J].J Exp Med,2004,200(2):267-272.
[18] Benard EL,Roobol SJ,Spaink HP,et al.Phagocytosis of mycobacteria by zebrafish macrophages is dependent on the scavenger receptor Marco,a key control factor of pro-inflammatory signalling[J].Dev Comp Immunol,2014,47(2):223-233.
[19] Li Z,Jiao Y,Fan EK,et al.Aging-impaired filamentous actin polymerization signaling reduces alveolar macrophage phagocytosis of bacteria[J].J Immunol,2017,199(9):3176-3186.