壳聚糖和普朗尼克介导猪白细胞介素-23基因体内表达对PRRSV疫苗的免疫增强效应
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摘要
猪繁殖和呼吸综合征(PRRS)是一种严重影响养猪业的高度传染性疾病,目前预防这种疾病的疫苗免疫还存在一定缺陷。本研究旨在探索一种有效的佐剂以提高其免疫效力。将猪IL-23基因重组质粒用壳聚糖和普朗尼克材料包裹制成纳米颗粒,命名为VRIL23-CNP-Pluronic。将28日龄小鼠分为两组,分别肌肉注射VRIL23-CNP-Pluronic(实验组)和VR1020-Pluronic(对照组),两组均同时接种PRRSV疫苗,在接种后第0、7、14、28和35天采集血样并分析免疫变化。实验组中PRRSV特异性抗体、IgG1和IgG2a水平、CD4+和CD8+T淋巴细胞数量均极显著高于对照组(p<0.01);经qRT-PCR分析,与对照组相比,实验组小鼠的IL-23、TLR1、TLR6、STAT1、IL-10、TNF-α、IL-15和CD62L基因表达水平均极显著上调(p<0.01)。结果表明,VRIL23-CNP-Pluronic能促进小鼠对PRRSV疫苗的免疫应答,并为其作为新型PRRSV疫苗佐剂的开发提供理论基础。
Porcine reproductive and respiratory syndrome(PRRS)is a severe,economically important infectious disease of pigs.Current vaccines to control the disease,however,are inadequate due to low immunity;and the present study was conducted to explore an effective adjuvant to enhance the immunological efficacy.A recombinant plasmid containing the porcine IL-23 gene was encapsulated in chitosan nanoparticles and Pluronic-L64 designated as VRIL23-CNP-Pluronic,which was tested following intramuscular injection of 28 day-old mice in comparison with VR1020-Pluronic as the control.Over a 35 day observation period,PRRSV-specific antibody,IgG1 and IgG2 a titers,CD4~+ and CD8~+ T cells increased significantly in the treated mice(P<0.01).As analyzed by qRT-PCR,expression levels of IL-23,TLR1,TLR6,STAT1,IL-10,TNF-α,IL-15 and CD62 L genes were also significantly up-regulated in comparison with those of control mice(P<0.01).These results show that co-injection of VRIL23-CNP-Pluronic with PRRS vaccine enhances both innate and adaptive immunity of mice,and provide support for its development as a novel vaccine adjuvant for control of swine PRRS.
Porcine reproductive and respiratory syndrome(PRRS)is a severe,economically important infectious disease of pigs.Current vaccines to control the disease,however,are inadequate due to low immunity;and the present study was conducted to explore an effective adjuvant to enhance the immunological efficacy.A recombinant plasmid containing the porcine IL-23 gene was encapsulated in chitosan nanoparticles and Pluronic-L64 designated as VRIL23-CNP-Pluronic,which was tested following intramuscular injection of 28 day-old mice in comparison with VR1020-Pluronic as the control.Over a 35 day observation period,PRRSV-specific antibody,IgG1 and IgG2 a titers,CD4~+ and CD8~+ T cells increased significantly in the treated mice(P<0.01).As analyzed by qRT-PCR,expression levels of IL-23,TLR1,TLR6,STAT1,IL-10,TNF-α,IL-15 and CD62 L genes were also significantly up-regulated in comparison with those of control mice(P<0.01).These results show that co-injection of VRIL23-CNP-Pluronic with PRRS vaccine enhances both innate and adaptive immunity of mice,and provide support for its development as a novel vaccine adjuvant for control of swine PRRS.
引文
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[21]Lin H W,Thompson J W,Morris K C,et al.Signal transducers and activators of transcription:STATs-mediated mitochondrial neuroprotection[J].Anti Redox Signal,2011,14(10):1 853-1 861.
[22]Lieberman L A,Zeng W,Singh C,et al.CD62L is not a reliable biomarker for predicting PML risk in natalizumab-treated R-MS patients[J].Neurology,2016,86(4):375-381.
[2]Lunney J K,Fang Y,Ladinig A,et al.Porcine reproductive and respiratory syndrome virus(prrsv):pathogenesis and interaction with the immune system[J].Annu Rev Anim Biosci,2016,4(1):129-54.
[3]Nan Y,Wu C,Gu G,et al.Improved vaccine against prrsv:current progress and future perspective[J].Front in Microbiol,2017,8:1 635-1 641.
[4]Jantafong T,Sangtong P,Saenglub W,et al.Genetic diversity of porcine reproductive and respiratory syndrome virus in Thailand and Southeast Asia from 2008 to 2013[J].Vet Microbiol,2015,176(3-4):229-238.
[5]Kang Y,Jin H,Zheng G,et al.The adjuvant effect of levamisole on killed viral vaccines[J].Vaccine,2005,23(48-49):5 543-5 550.
[6]Zengyu Z,Yaohong Z,Panqi L,et al.Effects of astragalus polysaccharide on immune responses of porcine pbmc stimulated with prrsv or csfv[J].Plos One,2012,7(1):S378-S379.
[7]Yijun D,Yu L,Xinglong W,et al.Highly efficient expression of interleukin-2 under the control of rabbit β-globin intron II gene enhances protective immune responses of porcine reproductive and respiratory syndrome(prrs)DNA vaccine in pigs[J].Plos One,2014,9(3):e90 326.
[8]Huabing Z,Xiaoping W,Guangming B,et al.Improvement of the immunity of piglets to prrs vaccine by a porcine IL-4 and IL-6 fusion gene encapsulated in chitosan nanoparticles[J].Procedia in Vaccinology,2012,6(6):113-124.
[9]陈祎,宋婷玉,李金海,等.壳聚糖纳米颗粒包裹猪白细胞介素2和融合白细胞介素4/6基因对猪圆环病毒2型疫苗免疫的强化[J].四川动物,2018,37(2):156-163.
[10]Oppmann B,Lesley R,Blom B,et al.Novel p19 protein engages IL-12p40 to form a cytokine,IL-23,with biological activities similar as well as distinct from IL-12[J].Immunity,2000,13(5):715-25.
[11]Langrish C L,Mckenzie B S,Wilson N J,et al.IL-12 and IL-23:master regulators of innate and adaptive immunity[J].Immunolo Rev,2010,202(1):96-105.
[12]Chamilos G,Ganguly D,Lande R,et al.Generation of IL-23 producing dendritic cells(DCs)by airborne fungi regulates fungal pathogenicity via the induction of TH-17 responses[J].Plos One,2010,5(9):e12955.
[13]Qiang X,Yonggang Z,Yanjun Z,et al.Immune responses of swine following DNA immunization with plasmids encoding porcine reproductive and respiratory syndrome virus ORFs 5 and 7,and porcine IL-2 and IFNγ[J].Vet Immunol Immunopathol,2004,102(3):291-298.
[14]Renukaradhya G J,Konstantin A,Kwonil J,et al.Porcine reproductive and respiratory syndrome virus-induced immunosuppression exacerbates the inflammatory response to porcine respiratory coronavirus in pigs[J].Viral Immunol,2010,23(5):457-466.
[15]Shin Foong N,Teng M W L,Smyth M J.A balance of interleukin-12 and-23 in cancer[J].Trends in Immunol,2013,34(11):548-555.
[16]Lopez O J,Osorio F A.Role of neutralizing antibodies in prrsv protective immunity[J].Vet Immunol Immunopathol,2004,102(3):155-163.
[17]Kabanov A V,Batrakova E V,Alakhov V Y.Pluronic? block copolymers as novel polymer therapeutics for drug and gene delivery[J].J Control Release,2002,82(2):189-212.
[18]Jianlin C,Jing L,Ying Z,et al.Increase in transgene expression by pluronic L64-mediated endosomal/lysosomal escape through its membrane-disturbing action[J].Acs Appl Mater Interfaces,2015,7(13):7 282-7 293.
[19]Choi J H,Jang J Y,Joung Y K,et al.Intracellular delivery and anti-cancer effect of self-assembled heparin-pluronic nanogels with RNase A[J].J Control Release,2010,147(3):420-427.
[20]Kawai T,Akira S.The role of pattern-recognition receptors in innate immunity:update on toll-like receptors[J].Nat Immunol,2010,11(5):373-384.
[21]Lin H W,Thompson J W,Morris K C,et al.Signal transducers and activators of transcription:STATs-mediated mitochondrial neuroprotection[J].Anti Redox Signal,2011,14(10):1 853-1 861.
[22]Lieberman L A,Zeng W,Singh C,et al.CD62L is not a reliable biomarker for predicting PML risk in natalizumab-treated R-MS patients[J].Neurology,2016,86(4):375-381.