酵母诱导表达鸡IFN-α抗IBDV效果及其对信号分子PI3K和NF-κB的影响

doi:10.16431/j.cnki.1671-7236.2016.08.032

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (8): 2164-2169.doi: 10.16431/j.cnki.1671-7236.2016.08.032

酵母诱导表达鸡IFN-α抗IBDV效果及其对信号分子PI3K和NF-κB的影响

韩海锋¹, 李银聚¹, 程相朝¹, 张春杰¹, 余祖华¹, 杨惠¹, 杨荣恩², 付威²

1. 河南科技大学动物疫病与公共安全重点实验室, 洛阳 471003;
2. 河南天纳图生物科技有限公司, 漯河 462600

收稿日期:2015-12-28 出版日期:2016-08-20 发布日期:2016-08-23
通讯作者: 李银聚 E-mail:lyj6511@126.com
作者简介:韩海锋(1989-),男,河南洛阳人,硕士生,研究方向:病毒分子生物学,E-mail:haifeng_han@139.com
基金资助:
河南省科技攻关项目（132102110117）

Study on the Anti-IBDV Activity of Chicken IFN-α and Its Effects on Signaling Molecule PI3K and NF-κB

HAN Hai-feng¹, LI Yin-ju¹, CHENG Xiang-chao¹, ZHANG Chun-jie¹, YU Zu-hua¹, YANG Hui¹, YANG Rong-en², FU Wei²

1. Key Laboratory of Animal Disease and Public Security, Henan University of Science and Technology, Luoyang 471003, China;
2. Henan Tian Na Tu Biotechnology Co., Ltd., Luohe 462600, China

Received:2015-12-28 Online:2016-08-20 Published:2016-08-23

摘要/Abstract

摘要：

试验旨在研究酵母表达鸡IFN-α抗传染性法氏囊病病毒（IBDV）的效果及对其淋巴细胞信号分子PI3K和NF-κB p65的影响。将40只10日龄非免疫雏鸡随机分为IFN-α组和空白对照组，持续注射给药3 d后，于第3次给药后第1天开始每天心脏采血，持续3 d，并于最后一次采血后对雏鸡进行攻毒，在攻毒后开始每天心脏采血，连续3 d。通过MTT法检测淋巴增殖活性情况，ELISA测定不同时间段淋巴细胞中PI3K的水平、细胞中总NF-κB p65、细胞核中NF-κB p65蛋白表达含量及IBDV抗原量。结果显示，与对照组相比，攻毒前，IFN-α对淋巴增殖活性、细胞中PI3K和NF-κB p65的表达具有极显著地促进作用（P<0.01）；与攻毒前相比，攻毒后第1和2天IFN-α组淋巴增殖活性、细胞中PI3K和NF-κB p65的表达极显著增加（P<0.01），IFN-α对IBDV-Ag具有极显著地抑制效果（P<0.01）。这些结果表明，IFN-α可通过增加PI3K激活NF-κB信号通路以增强机体免疫反应，并对IBDV-Ag有显著的抑制作用，本试验为IFN-α免疫增强和抗病毒作用的研究提供科学依据。

关键词: IFN-α PI3K; 核转录因子; NF-κ B p65; 传染性法氏囊病

Abstract:

To investigate the anti-IBDV activity of IFN-α and its effects on chicken lymphocyte signaling molecule PI3K and NF-κB activity,forty 10-day-old non-immune chickens were randomly separated into two groups:Blank control group and IFN-α group.The chicken of two groups were inoculated with IFN-α and saline via intramscular injection for three days,respectively.Then they were inoculated with IBDV.From 13 to 18-day-old,the lymphocytes were isolated from peripheral blood of chicken.The lymphocyte proliferation of chicken were detected by the MTT method,the PI3K and NF-κB p65 concentration in lymphocyte and the IBDV-Ag of serum were detected at different time with ELISA.The results showed that compared with the blank control group,IFN-α could stimulate chicken lymphocyte proliferation and the PI3K and NF-κB p65 concentration in chicken lymphocyte were extremely significantly increased before infection (P<0.01).The lymphocyte proliferation and the expression of PI3K and NF-κB p65 in cell after IBDV infection were extremely significantly higher than before (P<0.01).IFN-α could extremely significantly inhabit the proliferation of IBDV-Ag (P<0.01).IFN-α could significantly promote lymphocyte transformation by increasing PI3K concentration and the biological activity of lymphocyte transformation associated with the PI3K and NF-κB p65 expression,activation and nucleo-cytoplasmic transport.It also could reduce IBDV-Ag proliferation in chickens.The study built foundation for IFN-α protein immuno enhancement and anti-viral activity.

Key words: IFN-α; PI3K; nuclear factor; NF-κB p65; IBD

中图分类号:

S858.31

韩海锋, 李银聚, 程相朝, 张春杰, 余祖华, 杨惠, 杨荣恩, 付威. 酵母诱导表达鸡IFN-α抗IBDV效果及其对信号分子PI3K和NF-κB的影响[J]. 《中国畜牧兽医》, 2016, 43(8): 2164-2169.

HAN Hai-feng, LI Yin-ju, CHENG Xiang-chao, ZHANG Chun-jie, YU Zu-hua, YANG Hui, YANG Rong-en, FU Wei. Study on the Anti-IBDV Activity of Chicken IFN-α and Its Effects on Signaling Molecule PI3K and NF-κB[J]. , 2016, 43(8): 2164-2169.

参考文献

[1] Muller H,Islam M R,Raue R.Research on infectious bursal disease——The past,the present and the future[J].Veterinary Microbiology,2003,97(1-2):153-165.
[2] Wang Y S,Wang Z C,Tang Y D,et al.Comparison of four infectious bursal disease viruses isolated from different bird species[J].Archives of Virology,2007,152(10):1787-1797.
[3] 刘成倩,王静,李红,等.鸡干扰素-α基因原核表达载体的构建及高效表达[J].中国畜牧兽医,2014,41(4):47-50.
[4] Parmar S,Platanias L C.Interferons:Mechanisms of action and clinical applications[J].Current Opinion in Oncology,2003,15(6):431-439.
[5] Jensen M,Tawadros S,Sedlacek H H,et al.NK cell depletion diminish tumour-specific B cell responses[J].Immunology Letters,2004,93(2):205-210.
[6] Meager A.Biological assays for interferons[J].Journal of Immunological Methods,2002,261(1):21-36.
[7] Samuel C E.Antiviral actions of interferons[J].Clinical Microbiology Reviews,2001,14(4):778-809.
[8] 杨惠,李银聚,刘一尘,等.重组IFN-α-IL-18体外对鸡外周血淋巴细胞转化及核因子κB的影响[J].中国免疫学杂志,2015,31(8):1040-1044.
[9] Li Y J,Wang C.Synonymous codon usage of the VP2 gene of a very virulent infectious bursal disease virus isolate serial passaged in chicken embryos[J].Biosystems,2011,104:42-47.
[10] Helfand S C,Modiano J F,Nowell P C.Immunophysiological studies of interleukin-2 and canine lymphocytes[J].Veterinary Immunology and Immunopathology,1992,33(1):1-16.
[11] Tayade C,Koti M,Mishra S C.L-Arginine stimulates intestinal intraepithelial lymphocyte functions and immune response in chickens orally immunized with live intermediate plus strain of infectious bursal disease vaccine[J].Vaccine,2006,24(26):5473-5480.
[12] 孙静静,邵军军,常惠芸.干扰素作用机制及其在抗口蹄疫病毒中的应用研究进展[J].中国畜牧兽医,2012,39(1):206-209.
[13] Seidelin J B,Jaattela M,Nielsen O H.Continuous interferon-gamma or tumor necrosis factor-alpha exposure of enterocytes attenuates cell death responses[J].Cytokine,2004,27(4):113-119.
[14] Ferrantini M,Capone I,Belardelli F.Interferon-alpha and cancer:Mechanisms of action and new perspectives of clinical use[J].Biochimie,2007,89(6):884-893.
[15] Raison C L,Borisov A S,Woolwine B J,et al.Interferon-alpha effects on diurnal hypothalamic-pituitary-adrenal axis activity:Relationship with proinflammatory cytokines and behavior[J].Molecular Psychiatry,2010,15(5):535-547.
[16] Park J H,Sung H W,Yoon B I,et al.Protection of chicken against very virulent IBDV provided by in ovo priming with DNA vaccine and boosting with killed vaccine and the adjuvant effects of plasmid-encoded chicken interleukin-2 and interferon-gamma[J].Journal of Veterinary Science,2009,10(2):131-139.
[17] 张英楠,刘洪章,刘树英,等.免疫器官中淋巴细胞分布的研究进展[J].中国畜牧兽医,2014,41(2):124-128.
[18] Wu Q L,Shen T,Ma H,et al.Sufentanil postconditioning protects the myocardium from ischemia-reperfusion via PI3K/Akt-GSK-3beta pathway[J].The Journal of Surgical Research,2012,178(2):563-570.
[19] Wei L,Hou L,Zhu S,et al.Infectious bursal disease virus activates the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway by interaction of VP5 protein with the p85alpha subunit of PI3K[J].Virology,2011,417(1):211-220.
[20] Garcia M G,Alaniz L D,Cordo Russo R I,et al.PI3K/Akt inhibition modulates multidrug resistance and activates NF-kappaB in murine lymphoma cell lines[J].Leukemia Research,2009,33(2):288-296.
[21] Hawkins P T,Stephens L R.PI3K signalling in inflammation[J].Biochimicaet Biophysicaacta,2015,1851(6):882-897.

酵母诱导表达鸡IFN-α抗IBDV效果及其对信号分子PI3K和NF-κB的影响

Study on the Anti-IBDV Activity of Chicken IFN-α and Its Effects on Signaling Molecule PI3K and NF-κB

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	何献铭, 任广彩, 叶俊贤, 兰虹, 刘郁夫, 熊挺, 杨泽坤, 徐婷, 陈瑞爱. 1株传染性法氏囊病病毒的分离鉴定及VP2基因序列分析[J]. 中国畜牧兽医, 2022, 49(3): 1005-1014.
[2]	徐婷, 熊挺, 李淋雨, 刘郁夫, 温良海, 谢文婷, 杨泽坤, 陈瑞爱. 表达传染性法氏囊病病毒VP2蛋白的重组复制缺陷型HAdV-5的构建及鉴定[J]. 中国畜牧兽医, 2022, 49(12): 4524-4534.
[3]	陈欣, 达赖, 付绍印, 祁云霞, 王标, 何小龙, 马跃军, 方勤圆, 张林, 贾明蕊, 董鹏, 刘永斌. 绵羊卵泡发育过程中关键信号通路研究进展[J]. 中国畜牧兽医, 2021, 48(3): 963-974.
[4]	孟维金, 范阔海, 常熊熊, 尹伟, 李宏全, 王志瑞. 鸡传染性法氏囊病病毒VP2与白喉毒素截短体DT390融合蛋白的真核表达及免疫原性检测[J]. 中国畜牧兽医, 2021, 48(12): 4652-4660.
[5]	王彦伟, 张素玲, 吴芃, 耿笑林, 黄玉欣, 李鹏昊, 王孟月, 李向东, 逄文强, 田克恭. 鸡传染性法氏囊病病毒rVP2蛋白高效组装亚病毒颗粒的鉴定[J]. 中国畜牧兽医, 2020, 47(6): 1677-1684.
[6]	宋天琪, 侯绍华, 郭晓宇, 鑫婷, 姜一曈, 袁维峰, 朱鸿飞, 贾红. 重组犬α6干扰素的酵母表达及其抗病毒活性评价[J]. 《中国畜牧兽医》, 2019, 46(7): 2144-2150.
[7]	沈佳, 史爱华, 章振华, 李林, 景小冬, 张建伟, 郑小兰, 姜北宇. 鸡新城疫－传染性支气管炎－传染性法氏囊病三联灭活疫苗对不同日龄雏鸡的免疫效力研究[J]. 《中国畜牧兽医》, 2019, 46(3): 898-904.
[8]	朱艳平, 何勇, 刘佳, 邢瑞林, 常乐凯, 李润芷, 岳锋, 吴玉苹, 李鹏, 孙国鹏, 张艳芳, 王选年. 鸡传染性法氏囊病病毒VP2蛋白与幽门螺杆菌铁蛋白基因融合PCR扩增及其真核表达载体的构建[J]. 中国畜牧兽医, 2019, 46(10): 2860-2866.
[9]	高祥, 李辉, 张礼洲, 卢珍, 王永强, 高立, 吴甜甜, 高玉龙, 刘长军, 王笑梅, 祁小乐. 传染性法氏囊病病毒超强毒株衣壳蛋白的可溶性表达、纯化与活性分析[J]. 《中国畜牧兽医》, 2017, 44(7): 2096-2102.
[10]	郑华斌, 张中旺, 吕建亮, 潘丽, 张永光. 口蹄疫病毒多基因及猪α干扰素共表达载体的构建及其免疫豚鼠效果研究[J]. 《中国畜牧兽医》, 2017, 44(5): 1452-1461.
[11]	王洁琼, 赵玉杰, 周云飞, 黄宗梅, 陈盼盼, 周薇帆, 刘琳, 李新生. 传染性法氏囊病病毒C4毒株VP2基因的原核表达及生物信息学分析[J]. 《中国畜牧兽医》, 2017, 44(12): 3585-3591.
[12]	章振华, 李林, 景小冬, 张建伟, 沈佳, 史爱华, 郑小兰, 黄凤军, 姜北宇. 鸡传染性法氏囊病病毒BJQ902株在DF-1细胞系上增殖工艺研究[J]. 《中国畜牧兽医》, 2016, 43(10): 2775-2779.
[13]	章振华, 李林, 景小冬, 张建伟, 沈佳, 史爱华, 郑小兰, 黄凤军, 姜北宇. 鸡传染性法氏囊病免疫复合物疫苗对1日龄低母源抗体水平雏鸡免疫效果的研究[J]. 《中国畜牧兽医》---唯一指定的官方网站, 2015, 42(9): 2493-2498.
[14]	王念, 陈玉明, 张礼洲, 高立, 卢珍, 高玉龙, 王永强, 高宏雷, 刘长军, 崔红玉, 王笑梅, 祁小乐. 传染性法氏囊病病毒VP4酵母双杂交诱饵载体的构建及互作宿主细胞蛋白的筛选[J]. , 2015, 42(2): 317-323.
[15]	章振华, 李林, 景小冬, 张建伟, 沈佳, 史爱华, 郑小兰, 黄凤军, 姜北宇. 鸡传染性法氏囊病免疫复合物疫苗对1日龄中等母源抗体水平雏鸡免疫效果的研究[J]. , 2015, 42(12): 3338-3343.