噬菌体文库与计算机模拟筛选H1N1流感病毒血凝素结合位点的比较
|
摘要
目的:对H1N1流感病毒血凝素HA的抗原表位初步筛选。方法:用噬菌体展示文库对2株抗体的结合位点进行筛选,将筛选的抗原位点结合HA序列进行多肽合成,免疫小鼠制备单克隆抗体,通过分子生物学手段对4株抗体的轻链和重链可变区基因进行调取,通过计算机模拟抗体序列结合的抗原的结构域,推测抗体结合的氨基酸位点。结果:噬菌体文库筛选得出抗体的结合序列富含组氨酸和精氨酸,将针对多肽的单克隆抗体的序列和前期获得的2株抗HA单克隆抗体的序列比对分析,发现A1-8和anti-14p结合的HA抗原序列相一致,H1-13和anti-11p结合的HA抗原序列相一致。结论:通过将噬菌体展示文库技术和计算机模拟预测抗原抗体结合,可以有效筛选抗原表位,为流感病毒HA抗原表位预测方法的完善提供理论基础。
Objective: To screen the antigenic epitopes of hemagglutinin of H1 N1 influenza virus. Methods: The binding sites of the two antibodies with HA were analyzed by phage display library. Polypeptide synthesis based on the selected antigenic sites and HA sequence,monoclonal antibodies prepared by immunization mice with two polypeptide,light chain and heavy chain variable region of four antibodies were obtained by means of molecular cloning. Moreover,the binding domain of antibodies and antigen was predicted by computer simulation. Results: The phage library showed that the binding domain was a sequence of amino acids rich in histidine and arginine. The sequence of HA antigen binding to A1-8 and anti-14 p was consistent,and the sequence of HA antigen binding to H1-13 and anti-11 p was consistent. Conclusion: Predict antigen-antibody binding sites by the combination of phage display library technology and computer simulation may effectively screen antigen epitopes. This study may provide a theoretical basis for the improvement of influenza antigen HA epitope prediction.
Objective: To screen the antigenic epitopes of hemagglutinin of H1 N1 influenza virus. Methods: The binding sites of the two antibodies with HA were analyzed by phage display library. Polypeptide synthesis based on the selected antigenic sites and HA sequence,monoclonal antibodies prepared by immunization mice with two polypeptide,light chain and heavy chain variable region of four antibodies were obtained by means of molecular cloning. Moreover,the binding domain of antibodies and antigen was predicted by computer simulation. Results: The phage library showed that the binding domain was a sequence of amino acids rich in histidine and arginine. The sequence of HA antigen binding to A1-8 and anti-14 p was consistent,and the sequence of HA antigen binding to H1-13 and anti-11 p was consistent. Conclusion: Predict antigen-antibody binding sites by the combination of phage display library technology and computer simulation may effectively screen antigen epitopes. This study may provide a theoretical basis for the improvement of influenza antigen HA epitope prediction.
引文
[1]Huang SS,Banner D,Fang Y,et al.Comparative analyses of pandemic H1N1 and seasonal H1N1,H3N2,and influenza B infections depict distinct clinical pictures in ferrets[J].PLo S One,2011,6(11):1-14.
[2]Maines TR,Jayaraman A,Belser JA,et al.Transmission and pathogenesis of swine-origin 2009 A(H1N1)influenza viruses in ferrets and mice[J].Science,2009,325(5939):484-487.
[3]Nelson MI,Viboud C,Simonsen L,et al.Multiple reassortment events in the evolutionary history of H1N1 influenza A virus since1918[J].PLo S Pathog,2008,4:e1000012.
[4]Sanina N,Davydova L,Chopenko N,et al.Modulation of immunogenicity and conformation of HA1 subunit of influenza A virus H1/N1 hemagglutinin in tubular immunostimulating complexes[J].Int J Mol Sci,2017,18(9):E1895.
[5]Lansbury LE,Smith S,Beyer W,et al.Effectiveness of 2009 pandemic influenza A(H1N1)vaccines:A systematic review and meta-analysis[J].Vaccine,2017,35(16):1996-2006.
[6]Liu M,Zhao X,Hua S,et al.Antigenic patterns and evolution of the human influenza A(H1N1)virus[J].Sci Rep,2015,5:14171.
[7]Zhu W,Wang C,Wang BZ.From variation of influenza viral proteins to vaccine development[J].Int J Mol Sci,2017,18(7):E1554.
[8]蔡炯,钟彦伟,荆霞,等.噬菌体展示技术筛选甲型H1N1流感病毒抗原模拟表位的研究[J].解放军医学杂志,2010,35(1):61-63.Cai J,Zhong YW,Jing X,et al.Screening of mimotopes of swine influenza virus A(H1N1)by phage display technology[J].Med JChin PLA,2010,35(1):61-63.
[9]Cunha-Neto E,Rosa DS,Harris PE,et al.An approach for a synthetic CTL vaccine design against zika flavivirus using class I and class II epitopes identified by computer modeling[J].Front Immunol,2017,8:640.
[10]邱宇,曹丽,郝晓宁,等.肺癌T7噬菌体文库中MUC1蛋白抗原模拟表位的筛选[J].免疫学杂志,2016,32(2):132-136.Qiu Y,Cao L,Hao XN,et al.Screening of MUC1 mimotope in T7phage library of lung cancer[J].Immunol J,2016,32(2):132-136.
[11]金伯泉.细胞和分子免疫学实验技术[M].西安:第四军医大学出版社,2002:45-46.Jin BQ.Cell and molecular immunology experimental techniques[M].Xi'an:Fourth Military Medical University Press,2002:45-46.
[12]邓颖琦,李俊峰,曲辉,等.表达H1N1亚型猪流感病毒三聚体HA重组慢病毒的免疫原性研究[J].中国免疫学杂志,2016,32(7):983-987.Deng YQ,Li JF,Qu H,et al.Study on immunogenicity of recombinant lentivirus expressing trimeric HA of swine H1N1 influenza virus[J].Chin J Immunol,2016,32(7):983-987.
[13]李慧瑾,郭春艳,赵彭花,等.运用单克隆抗体分析流感病毒H1亚型HA蛋白的抗原表位[J].西安交通大学学报(医学版),2012,33(3):291-294.Li HJ,Guo CY,Zhao PH,et al.Analysis of epitopes on HA of H1subtype influenza virus using monoclonal antibodies[J].J Xi'an Jiaotong Univ(Med Sci),2012,33(3):291-294.
[14]Ma C,Pan Y,Zhang L,et al.Influenza vaccine effectiveness against medically attended influenza illness in Beijing,China,2014/15 season[J].Hum Vaccin Immunother,2017,13(10):2379-2384.
[15]Gao JX,Zhao YL,Liu JF,et al.Safety and effectiveness assessment of 2011-2012 seasonal influenza vaccine produced in China:a randomized trial[J].Postgrad Med,2017,129(8):907-914.
[16]Baxter D.Evaluating the case for trivalent or quadrivalent influenza vaccines[J].Hum Vaccin Immunother,2016,12(10):2712-2717.
[17]Duffy J,Lewis M,Harrington T,et al.Live attenuated influenza vaccine use and safety in children and adults with asthma[J].Ann Allergy Asthma Immunol,2017,118(4):439-444.
[18]Johansen K,Brasseur D,Mac Donald N,et al.Where are we in our understanding of the association between narcolepsy and one of the 2009 adjuvanted influenza A(H1N1)vaccines?[J].Biologicals,2016,44(4):276-280.
[19]赵奇,王远强,林勇,等.H5N1型高致病性禽流感病毒血凝素蛋白及神经氨酸酶的B细胞抗原表位预测[J].免疫学杂志,2010,26(5):408-411.Zhao Q,Wang YQ,Lin Y,et al.Prediction of the common B cell epitopes derived from hemagglutinin and neuraminidase of highly pathogenic avian influenza H5N1[J].Immunol J,2010,26(5):408-411.
[20]Cui Y,Yu L,Zhou Y,et al.Mimotope identification of dust mite allergen Der f 5 using phage displayed random peptide libraries[J].Mol Med Rep,2016,14(5):4816-4822.
[21]Riemer AB,Kraml G,Scheiner O,et al.Matching of trastuzumab(Herceptin)epitope mimics onto the surface of Her-2/neu--a new method of epitope definition[J].Mol Immunol,2005,42(9):1121-1124.
[22]Masoodi TA,Shaik NA,Shafi G,et al.Comparative analysis of hemagglutinin of 2009 H1N1 influenza A pandemic indicates its evolution to 1918 H1N1 pandemic[J].Gene,2012,491(2):200-204.
[2]Maines TR,Jayaraman A,Belser JA,et al.Transmission and pathogenesis of swine-origin 2009 A(H1N1)influenza viruses in ferrets and mice[J].Science,2009,325(5939):484-487.
[3]Nelson MI,Viboud C,Simonsen L,et al.Multiple reassortment events in the evolutionary history of H1N1 influenza A virus since1918[J].PLo S Pathog,2008,4:e1000012.
[4]Sanina N,Davydova L,Chopenko N,et al.Modulation of immunogenicity and conformation of HA1 subunit of influenza A virus H1/N1 hemagglutinin in tubular immunostimulating complexes[J].Int J Mol Sci,2017,18(9):E1895.
[5]Lansbury LE,Smith S,Beyer W,et al.Effectiveness of 2009 pandemic influenza A(H1N1)vaccines:A systematic review and meta-analysis[J].Vaccine,2017,35(16):1996-2006.
[6]Liu M,Zhao X,Hua S,et al.Antigenic patterns and evolution of the human influenza A(H1N1)virus[J].Sci Rep,2015,5:14171.
[7]Zhu W,Wang C,Wang BZ.From variation of influenza viral proteins to vaccine development[J].Int J Mol Sci,2017,18(7):E1554.
[8]蔡炯,钟彦伟,荆霞,等.噬菌体展示技术筛选甲型H1N1流感病毒抗原模拟表位的研究[J].解放军医学杂志,2010,35(1):61-63.Cai J,Zhong YW,Jing X,et al.Screening of mimotopes of swine influenza virus A(H1N1)by phage display technology[J].Med JChin PLA,2010,35(1):61-63.
[9]Cunha-Neto E,Rosa DS,Harris PE,et al.An approach for a synthetic CTL vaccine design against zika flavivirus using class I and class II epitopes identified by computer modeling[J].Front Immunol,2017,8:640.
[10]邱宇,曹丽,郝晓宁,等.肺癌T7噬菌体文库中MUC1蛋白抗原模拟表位的筛选[J].免疫学杂志,2016,32(2):132-136.Qiu Y,Cao L,Hao XN,et al.Screening of MUC1 mimotope in T7phage library of lung cancer[J].Immunol J,2016,32(2):132-136.
[11]金伯泉.细胞和分子免疫学实验技术[M].西安:第四军医大学出版社,2002:45-46.Jin BQ.Cell and molecular immunology experimental techniques[M].Xi'an:Fourth Military Medical University Press,2002:45-46.
[12]邓颖琦,李俊峰,曲辉,等.表达H1N1亚型猪流感病毒三聚体HA重组慢病毒的免疫原性研究[J].中国免疫学杂志,2016,32(7):983-987.Deng YQ,Li JF,Qu H,et al.Study on immunogenicity of recombinant lentivirus expressing trimeric HA of swine H1N1 influenza virus[J].Chin J Immunol,2016,32(7):983-987.
[13]李慧瑾,郭春艳,赵彭花,等.运用单克隆抗体分析流感病毒H1亚型HA蛋白的抗原表位[J].西安交通大学学报(医学版),2012,33(3):291-294.Li HJ,Guo CY,Zhao PH,et al.Analysis of epitopes on HA of H1subtype influenza virus using monoclonal antibodies[J].J Xi'an Jiaotong Univ(Med Sci),2012,33(3):291-294.
[14]Ma C,Pan Y,Zhang L,et al.Influenza vaccine effectiveness against medically attended influenza illness in Beijing,China,2014/15 season[J].Hum Vaccin Immunother,2017,13(10):2379-2384.
[15]Gao JX,Zhao YL,Liu JF,et al.Safety and effectiveness assessment of 2011-2012 seasonal influenza vaccine produced in China:a randomized trial[J].Postgrad Med,2017,129(8):907-914.
[16]Baxter D.Evaluating the case for trivalent or quadrivalent influenza vaccines[J].Hum Vaccin Immunother,2016,12(10):2712-2717.
[17]Duffy J,Lewis M,Harrington T,et al.Live attenuated influenza vaccine use and safety in children and adults with asthma[J].Ann Allergy Asthma Immunol,2017,118(4):439-444.
[18]Johansen K,Brasseur D,Mac Donald N,et al.Where are we in our understanding of the association between narcolepsy and one of the 2009 adjuvanted influenza A(H1N1)vaccines?[J].Biologicals,2016,44(4):276-280.
[19]赵奇,王远强,林勇,等.H5N1型高致病性禽流感病毒血凝素蛋白及神经氨酸酶的B细胞抗原表位预测[J].免疫学杂志,2010,26(5):408-411.Zhao Q,Wang YQ,Lin Y,et al.Prediction of the common B cell epitopes derived from hemagglutinin and neuraminidase of highly pathogenic avian influenza H5N1[J].Immunol J,2010,26(5):408-411.
[20]Cui Y,Yu L,Zhou Y,et al.Mimotope identification of dust mite allergen Der f 5 using phage displayed random peptide libraries[J].Mol Med Rep,2016,14(5):4816-4822.
[21]Riemer AB,Kraml G,Scheiner O,et al.Matching of trastuzumab(Herceptin)epitope mimics onto the surface of Her-2/neu--a new method of epitope definition[J].Mol Immunol,2005,42(9):1121-1124.
[22]Masoodi TA,Shaik NA,Shafi G,et al.Comparative analysis of hemagglutinin of 2009 H1N1 influenza A pandemic indicates its evolution to 1918 H1N1 pandemic[J].Gene,2012,491(2):200-204.