Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2017, Vol. 37 Issue (4): 26-32    DOI: 10.13523/j.cb.20170404
研究报告     
重组小鼠白细胞介素-33的原核表达制备及其粘膜免疫佐剂活性
冯雪军1,2,3, 龙琼1,2,3, 唐增华1,2,3, 黄惟巍1,2,3, 刘存宝1,2,3, 杨旭1,2,3, 孙文佳1,2,3, 白红妹1,2,3, 马雁冰1,2,3
1 中国医学科学院、北京协和医学院 医学生物学研究所 昆明 650118;
2 云南省重大传染病疫苗研发重点实验室 昆明 650118;
3. 云南省重大传染病疫苗工程技术研究中心 昆明 650118
The Prokaryotic Expression of Recombinant Murine Interlukin (IL)-33 and Its Mucosal Immunization Affect as Adjuvant
FENG Xue-jun1,2,3, LONG Qiong1,2,3, TANG Zeng-hua1,2,3, HUANG Wei-wei1,2,3, LIU Cun-bao1,2,3, YANG Xu1,2,3, SUN Wen-jia1,2,3, BAI Hong-mei1,2,3, MA Yan-bing1,2,3
1 Institute of Medical Biology, Chinese Academy of Medical Science & Peking Union Medical College, Kunming 650118, China;
2 Yunnan Key Laboratory of Vaccine Research & Developent on Severe Infection Disease, Kunming 650118, China;
3 Yunnan Engineering Research of Vaccine Research and Development on Severe Infection Disease, Kunming 650118, China
 全文: PDF(839 KB)   HTML
摘要: 目的:白细胞介素 (IL)-33对树突状细胞、巨噬细胞及T细胞等免疫细胞具有重要调控作用。利用大肠杆菌制备重组小鼠的IL-33,并初步考察其作为粘膜免疫佐剂应用的潜能与特点。方法:以IPTG诱导硫氧还蛋白(Trx)/IL-33融合蛋白在大肠杆菌DH5α中的表达,并通过Q-Sepharose离子交换和Ni++金属螯合亲和层析纯化Trx/IL-33,进一步经肠激酶切割获得成熟形式的IL-33。重组HBcAg混合纯化的IL-33后经滴鼻免疫小鼠,考察HBcAg特异的IgA及IgG1、IgG2a的应答。结果:纯化的重组IL-33具有与标准品相当的促巨噬细胞RAW264.7表达TNF-α的体外细胞生物学活性。作为佐剂可显著增强滴鼻粘膜免疫激发的不同粘膜组织中HBcAg特异的IgA应答,以及血清与支气管肺泡灌洗液中特异IgG1的应答水平,而抑制IgG2a应答。结论:利用大肠杆菌可制备活性IL-33,其具有粘膜免疫佐剂的应用潜能。
关键词: 白细胞介素-33佐剂粘膜免疫重组表达    
Abstract: Objective:Interleukin(IL)-33 has important immunoregulatory effects on immune cells, such as dentritic cells, macropphage and T cells. The purpose is to prepare recombinant murine IL-33 in E.coli cells, and investigate the potentials and characteristics of IL-33 to be employed as an adjuvant in mucosal immunization. Methods:The expression of fusion protein thioredoxin/IL-33 was induced by IPTG in DH5α cells, and the purification was performed through a chromatography procedure consists of Q-Sepharose ionic exchange and Ni++ chelating affinity. And then, mature IL-33 was released from the fusion protein by enterokinase treatment. The recombinant HBcAg was combined with IL-33 or not, and used to immunize mice intranasally. HBcAg-specific IgA, IgG1 and IgG2a were detected with ELISAs. Results:Purified IL-33 displayed the similar bioactivity of stimulating RAW264.7 to express TNF-α as commercially derived recombinant IL-33. Purified IL-33 which served as an adjuvant significantly enhanced the HBcAg-specific IgA response in multiple mucosal tissues, as well as IgG1 levels in serum and bronchial alveolar lavage fluid, whereas reduced IgG2a levels. Conclusion:Active IL-33 is successfully prepared from E.coli cells, and has a potential to be used as a molecular adjuvant in mucosal immunization.
Key words: Interlukin (IL)-33    Mucosal immunization    Adjuvant    Expression    Recombinant
收稿日期: 2016-11-01 出版日期: 2017-04-25
ZTFLH:  Q819  
基金资助: 云南省对外合作科技项目(2013IA005)、云南省应用基础研究重点项目(2011FA023,2016FA023)资助项目
通讯作者: 马雁冰     E-mail: may@imbcams.com.cn
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
冯雪军
唐增华
黄惟巍
杨旭
白红妹
马雁冰
孙文佳
龙琼
刘存宝

引用本文:

冯雪军, 龙琼, 唐增华, 黄惟巍, 刘存宝, 杨旭, 孙文佳, 白红妹, 马雁冰. 重组小鼠白细胞介素-33的原核表达制备及其粘膜免疫佐剂活性[J]. 中国生物工程杂志, 2017, 37(4): 26-32.

FENG Xue-jun, LONG Qiong, TANG Zeng-hua, HUANG Wei-wei, LIU Cun-bao, YANG Xu, SUN Wen-jia, BAI Hong-mei, MA Yan-bing. The Prokaryotic Expression of Recombinant Murine Interlukin (IL)-33 and Its Mucosal Immunization Affect as Adjuvant. China Biotechnology, 2017, 37(4): 26-32.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170404        https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I4/26

[1] Schmitz J, Owyang A, Oldham E, et al. IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity, 2005,23(5):479-490.
[2] Moussion C, Ortega N, Girard J P. The IL-1-like cytokine IL-33 is constitutively expressed in the nucleus of endothelial cells and epithelial cells in vivo:a novel ‘alarmin’? PLoS One, 2008,3(10):e3331.
[3] Komai-Koma M, Xu D, Li Y, et al. IL-33 is a chemoattractant for human Th2 cells. Eur J Immunol, 2007,37(10):2779-2786.
[4] Su Z, Lin J, Lu F, et al. Potential autocrine regulation of interleukin-33/ST2 signaling of dendritic cells in allergic inflammation. Mucosal Immunology, 2013,6(5):921-930.
[5] Luzina I G, Pickering E M, Kopach P, et al. Full-length IL-33 promotes inflammation but not Th2 response in vivo in an ST2-independent fashion. Journal of Immunology, 2012,189(1):403-410.
[6] Bourgeois E, Van L P, Samson M, et al. The pro-Th2 cytokine IL-33 directly interacts with invariant NKT and NK cells to induce IFN-gamma production. Eur J Immunol, 2009,39(4):1046-1055.
[7] Bonilla W V, Frohlich A, Senn K, et al. The alarmin interleukin-33 drives protective antiviral CD8+ T cell responses. Science, 2012,335(6071):984-989.
[8] Gao X, Wang X, Yang Q, et al. Tumoral expression of IL-33 inhibits tumor growth and modifies the tumor microenvironment through CD8+ T and NK cells. Journal of Immunology, 2015,194(1):438-445.
[9] Villarreal D O, Wise M C, Walters J N, et al. Alarmin IL-33 acts as an immunoadjuvant to enhance antigen-specific tumor immunity. Cancer Res, 2014,74(6):1789-1800.
[10] Froidure A, Vandenplas O, D'Alpaos V, et al. Persistence of asthma following allergen avoidance is associated with proTh2 myeloid dendritic cell activation. Thorax, 2015,70(10):967-973.
[11]. Chu Xiaojie,Li Yang,Long Qiong,et al.Development of a HPV16 E7 CTLs peptides-based virus-like particle therapertic vaccine.China Biotechnology,2015,35(2):45-51.
[12] Gao K, Li X, Zhang L, et al. Transgenic expression of IL-33 activates CD8(+) T cells and NK cells and inhibits tumor growth and metastasis in mice. Cancer Letters, 2013,335(2):463-471.
[13] Bagarazzi M L, Yan J, Morrow M P, et al. Immunotherapy against HPV16/18 generates potent TH1 and cytotoxic cellular immune responses. Science Translational Medicine, 2012,4(155):138.
[1] 贺立恒,张毅,张洁,任豫超,解红娥,唐锐敏,贾小云,武宗信. 基于转录组和WGCNA的甘薯花青素合成相关基因共表达网络的构建及核心基因的挖掘*[J]. 中国生物工程杂志, 2021, 41(9): 27-36.
[2] 乔圣泰,王曼琦,徐慧妮. 番茄SlTpx原核表达蛋白的体外功能分析*[J]. 中国生物工程杂志, 2021, 41(8): 25-32.
[3] 郭曼曼,田开仁,乔建军,李艳妮. 噬菌体重组酶系统在合成生物学中的应用*[J]. 中国生物工程杂志, 2021, 41(8): 90-102.
[4] 李冰,张传波,宋凯,卢文玉. 生物合成稀有人参皂苷的研究进展*[J]. 中国生物工程杂志, 2021, 41(6): 71-88.
[5] 黄蕾,万常青,刘美琴,赵敏,郑妍鹏,彭向雷,虞结梅,付远辉,何金生. 利用DNA Assembly方法构建重组腺病毒载体[J]. 中国生物工程杂志, 2021, 41(6): 23-26.
[6] 马巧妮,王萌,朱兴全. 重组酶介导扩增技术及其在病原微生物快速检测中的应用进展*[J]. 中国生物工程杂志, 2021, 41(6): 45-49.
[7] 张磊,唐永凯,李红霞,李建林,徐逾鑫,李迎宾,俞菊华. 促进原核表达蛋白可溶性的研究进展 *[J]. 中国生物工程杂志, 2021, 41(2/3): 138-149.
[8] 刘美琴,高博,焦月盈,李玮,虞结梅,彭向雷,郑妍鹏,付远辉,何金生. 人呼吸道合胞病毒感染的A549细胞中长链非编码RNA表达谱研究[J]. 中国生物工程杂志, 2021, 41(2/3): 7-13.
[9] 王惠临,周凯强,朱红雨,王力景,杨仲璠,徐明波,曹荣月. 凝血因子VII及其重组表达新进展[J]. 中国生物工程杂志, 2021, 41(2/3): 129-137.
[10] 杨茜,栾雨时. sly-miR399在番茄抗晚疫病中的初步探究*[J]. 中国生物工程杂志, 2021, 41(11): 23-31.
[11] 陈素芳,夏明印,曾丽艳,安晓琴,田敏芳,彭建. 抗菌肽Cec4a的重组表达和抗菌活性研究*[J]. 中国生物工程杂志, 2021, 41(10): 12-18.
[12] 朱潇静,王芮,张欣欣,靳家鑫,路闻龙,丁大顺,霍翠梅,李青梅,孙爱军,庄国庆. 利用细菌人工染色体技术构建整合F基因的重组MDV疫苗株*[J]. 中国生物工程杂志, 2021, 41(10): 33-41.
[13] 石鹏程, 纪晓俊. 酵母系统表达人表皮生长因子研究进展 *[J]. 中国生物工程杂志, 2021, 41(1): 72-79.
[14] 饶海密,梁冬梅,李伟国,乔建军,财音青格乐. 真菌芳香聚酮化合物的合成生物学研究进展*[J]. 中国生物工程杂志, 2020, 40(9): 52-61.
[15] 邓通,周海胜,吴坚平,杨立荣. 基于分子伴侣策略提高NADPH依赖型醇脱氢酶的异源可溶性表达 *[J]. 中国生物工程杂志, 2020, 40(8): 24-32.