人兽共患寄生虫病候选疫苗分子筛选方法研究进展

doi:10.16431/j.cnki.1671-7236.2021.06.031

摘要/Abstract

摘要： 人兽共患寄生虫种类多、宿主广泛且危害严重。血吸虫病、棘球蚴病、囊尾蚴病、旋毛虫病、弓形虫病等是常见的重要人兽共患寄生虫病。人类和家畜饱受寄生虫病的危害，这对公共卫生和畜牧业造成了很大的影响。控制传染源、切断传播途径和保护易感群是控制人兽共患寄生虫病流行的综合防控措施。在综合防控策略中，疫苗的使用是切断循环链、控制乃至消灭人兽共患寄生虫病的理想和有效途径之一。选用高效的抗原筛选方法挖掘潜在的疫苗候选分子是开发疫苗的前提和关键。抗原筛选技术的更新换代使得研究者发掘出了更多新抗原和保护性多肽。现有的抗原筛选方法主要包括传统的粗抗原筛选法、cDNA文库筛选法、蛋白质组学筛选法、生物信息学及多组学技术联合筛选法。很多抗原筛选的方法是伴随寄生虫疫苗研究的发展应运而生的，粗抗原筛选法是基于抗原抗体相互反应的免疫学原理而设计的，此方法筛选的天然抗原可引起机体较强的免疫反应；cDNA文库筛选抗原的优势在于筛选更有针对性，所以候选产物的成分更单一、明确；蛋白质组学筛选法是基于质谱而兴起的一种筛选技术，它既可对未知蛋白组分进行鉴定，还可对鉴定结果进行差异比较，在未知分子的发现和功能特殊的靶分子筛选中发挥着重要作用；随着后基因时代的到来，生物信息学及多组学联合筛选技术使得抗原筛选逐步进入了多维、立体的筛选模式，也使得候选抗原及其表位的功能研究更加深入，这为基因工程疫苗和多肽疫苗候选分子的筛选提供了技术手段。

关键词: 寄生虫; 疫苗; 抗原筛选; cDNA文库; 蛋白质组学; 生物信息学; 多组学

Abstract: There are many kinds of zoonotic parasites with a wide range of hosts and serious damage.Schistosomiasis,echinococcosis,cysticercosis,trichinosis and toxoplasmosis are common parasitic zoonoses.Humans and livestock suffer from parasitic diseases,which have a significant impact on public health and livestock farming.Controlling the source of infection,cutting off the route of transmission and protecting susceptible groups are the comprehensive prevention and control measures to control the epidemic of parasitic zoonoses.In particular,vaccinations is one of the ideal and effective ways to cut off parasites’ life cycle,and control and even eliminate parasitic zoonoses.It is the premise and key to develop a vaccine to select an effective antigen screening method to excavate the potential candidate molecules for vaccine.With the development of antigen screening technology,more new antigens and protective peptides have been discovered.Existing antigen screening methods mainly include the traditional crude antigen screening method,cDNA library screening method,proteomics screening method,bioinformatics and multi-omics technology screening method.Many methods of antigen screening were developed along with the process of parasite vaccine research.The crude antigen screening method is based on the immunological principle of antigen-antibody interaction,and natural antigen screened by this method can cause a strong immune response.The advantage of cDNA library screening is that the screening is more targeted,so the composition of the candidate product is more single and clear.Proteomic screening technique is based on mass spectrometry,which can not only identify the unknown proteins,but also compare the differences of the identification results.It plays an important role in the discovery of unknown molecules and the screening of target molecules with special functions.With the advent of the post-genetic era,the combined screening technology of bioinformatics and multi-omics makes antigen screening turn into a multi-dimensional screening mode,and also makes the functional studies of candidate antigens and their epitopes more in-depth,which provides a technical means for the screening of candidate molecules of genetically engineered vaccines and polypeptide vaccines.

Key words: parasite; vaccine; antigen screening; cDNA library; proteomics; bioinformatics; multi-omics

中图分类号:

S855.9

代国栋, 闫鸿斌, 李立, 付宝权, 贾万忠. 人兽共患寄生虫病候选疫苗分子筛选方法研究进展[J]. 中国畜牧兽医, 2021, 48(6): 2177-2187.

DAI Guodong, YAN Hongbin, LI Li, FU Baoquan, JIA Wanzhong. Research Progress on Screening Methods for Candidate Vaccine Molecules Against Parasitic Zoonoses[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(6): 2177-2187.

参考文献

[1] 黄达娜,张仁利,唐屹君,等.被忽视的热带病防控现状[J].新发传染病电子杂志,2019,4(3):177-180. HUANG D N,ZHANG R L,TANG Y J,et al.Status in the prevention and control of neglected tropical diseases[J].Electronic Journal of Emerging Infectious Diseases,2019,4(3):177-180.(in Chinese)
[2] 庞建达,王海英,杜立银,等.猪带绦虫囊尾蚴病特异性抗原筛选的研究进展[J].中国农学通报,2017,33(11):131-136. PANG J D,WANG H Y,DU L Y,et al.Research progress of specific antigen screening for taeniasis/cysticercosis[J].Chinese Agricultural Science Bulletin,2017,33(11):131-136.(in Chinese)
[3] 徐梦飞,卢平萍,马勋,等.细粒棘球绦虫蛋白质组学研究进展[J].畜牧兽医学报,2018,49(3):466-476. XU M F,LU P P,MA X,et al.Advances in Echinococcus granulosus proteomics[J].Acta Veterinaria et Zootechnica Sinica,2018,49(3):466-476.(in Chinese)
[4] LEROUX L P,NASR M,VALANPARAMBIL R,et al.Analysis of the Trichuris suis excretory/secretory proteins as a function of life cycle stage and their immunomodulatory properties[J].Scientific Reports,2018,8(1):15921.
[5] GOOD M F,STANISIC D I.Whole parasite vaccines for the asexual blood stages of Plasmodium[J].Immunological Reviews,2019,293(1):270-282.
[6] DARCY F,DESLEE D,SANTORO F,et al.Induction of a protective antibody-dependent response against toxoplasmosis by in vitro excreted/secreted antigens from tachyzoites of Toxoplasma gondii[J].Parasite Immunology,1988,10(5):553-567.
[7] RICKARD M D,ADOLPH A J.Vaccination of lambs against infection with Taenia ovis using antigens collected during short-term in vitro incubation of activated T.ovis oncospheres[J].Parasitology,1977,75(2):183-188.
[8] RAJASEKARIAH G R,MITCHELL G F,RICKARD M D.Taenia taeniaeformis in mice:Protective immunization with oncospheres and their products[J].International Journal for Parasitology,1980,10(2):155-160.
[9] BENITEZ L G T H.Cloning and sequencing of the gene encoding the principal 18 kDa secreted antigen of activated oncospheres of Taenia saginata[J].Molecular & Biochemical Parasitology,1996,78(1-2):265-268.
[10] HARRISON G B L H.Identification and cDNA cloning of two novel low molecular weight host-protective antigens from Taenia ovis oncospheres[J].International Journal for Parasitology,1996,26(2):195-204.
[11] KASPER L H,CRABB J H,PFEFFERKORN E R.Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody[J].Journal of Immunology,1983,130(5):2407-2412.
[12] 才学鹏,郑亚东,贾万忠,等.猪带绦虫六钩蚴45W基因家族分子克隆[J].中国农业科学,2005,38(10):2117-2123. CAI X P,ZHENG Y D,JIA W Z,et al.Molecular cloning of 45W gene family of Taenia solium oncosphere[J].Scientia Agricultura Sinica,2005,38(10):2117-2123.(in Chinese)
[13] GAUCI C,MERLI M,MULLER V,et al.Molecular cloning of a vaccine antigen against infection with the larval stage of Echinococcus multilocularis[J].Infection and Immunity,2002,70(7):3969-3972.
[14] LIGHTOWLERS M W,ROLFE R,GAUCI C G.Taenia saginata:Vaccination against cysticercosis in cattle with recombinant oncosphere antigens[J].Experimental Parasitology,1996,84(3):330-338.
[15] GAUCI C,VURAL G,ÖNCEL T,et al.Vaccination with recombinant oncosphere antigens reduces the susceptibility of sheep to infection with Taenia multiceps[J].International Journal for Parasitology,2008,38(8-9):1041-1050.
[16] 刘太峰,孟庆峰,段小波,等.cDNA文库构建方法的研究进展[J].中国畜牧兽医,2012,39(7):40-43. LIU T F,MENG Q F,DUAN X B,et al.Research progress of construction methods of cDNA library[J].China Animal Husbandry & Veterinary Medicine,2012,39(7):40-43.(in Chinese)
[17] 贾传礼.毒害艾美耳球虫配子体cDNA文库的构建与筛选及其动力蛋白基因的表达[D].扬州:扬州大学,2017. JIA C L.The construction and screen of cDNA library for the Eimeria necatrix gametocytes and expression of dynein gene[D].Yangzhou:Yangzhou University,2017.(in Chinese)
[18] 冉旭华,闻晓波,王春仁,等.肝片形吸虫重组组织蛋白酶L的免疫反应性和免疫原性分析[J].中国寄生虫学与寄生虫病杂志,2014,32(4):289-292. RAN X H,WEN X B,WANG C R,et al.Immunoreactivity and immunogenicity analysis of the recombinant cathepsin L-like protease of Fasciola hepatica in SD rats[J].Chinese Journal of Parasitology and Parasitic Diseases,2014,32(4):289-292.(in Chinese)
[19] JOHNSON K S,HARRISON G B L,LIGHTOWLERS M W,et al.Vaccination against ovine cysticercosis using a defined recombinant antigen[J].Nature,1989,338(6216):585-587.
[20] LIGHTOWLERS M W,LIU D Y,HARALAMBOUS A,et al.Subunit composition and specificity of the major cyst fluid antigens of Echinococcus granulosus[J].Molecular and Biochemical Parasitology,1989,37(2):171.
[21] WOOLLARD D J,GAUCI C G,HEATH D D,et al.Protection against hydatid disease induced with the EG95 vaccine is associated with conformational epitopes[J].Vaccine,2000,19(4-5):498-507.
[22] FU Y,MARTINEZ C,CHALAR C,et al.A new potent antigen from Echinococcus granulosus associated with muscles and tegument[J].Molecular and Biochemical Parasitology,1999,102(1):43-52.
[23] BURG J L,PERELMAN D,KASPER L H,et al.Molecular analysis of the gene encoding the major surface antigen of Toxoplasma gondii[J].Journal of Immunology,1988,141(10):3584-3591.
[24] ZHANG Y,TAYLOR M G,MCCROSSAN M V,et al.Molecular cloning and characterization of a novel Schistosoma japonicum "irradiated vaccine-specific" antigen,Sj14-3-3[J].Molecular and Biochemical Parasitology,1999,103(1):25-34.
[25] RASSY D,BOBES R J,ROSAS G,et al.Characteriza-tion of S3Pvac anti-cysticercosis vaccine components:Implications for the development of an anti-cestodiasis vaccine[J].PLoS One,2010,5(6):e11287.
[26] 孙树汉,王俊霞,陈蕊雯,等.囊虫病诊断用抗原编码CDNA的分子克隆[J].中国寄生虫学与寄生虫病杂志,1997,15(1):18-23. SUN S H,WANG J X,CHEN R W,et al.Molecular cloning of cDNA encoding immunodiagnostic antigens of cysticercosis[J].Chinese Journal of Parasitology and Parasitic Diseases,1997,15(1):18-23.(in Chinese)
[27] WANG Q,SUN S,HU Z,et al.Immune response and protection elicited by DNA immunisation against Taenia cysticercosis[J].Vaccine,2003,21(15):1672-1680.
[28] 王欣悦,张莉.蛋白质组学及蛋白质翻译后修饰在畜牧领域中的应用研究进展[J].中国畜牧兽医,2019,46(4):1063-1073. WANG X Y,ZHANG L.Research progress on application of proteomics and post-translational modification of proteins in animal husbandry[J].China Animal Husbandry & Veterinary Medicine,2019,46(4):1063-1073.(in Chinese)
[29] SUNDAR S,SINGH B.Understanding Leishmania parasites through proteomics and implications for the clinic[J].Expert Review of Proteomics 2018,15(5):371-390.
[30] ZHANG L,ELIAS J E.Relative protein quantification using tandem mass tag mass spectrometry[J].Methods in Molecular Biology,2017,1550:185-198.
[31] 李君良.细粒棘球绦虫原头蚴蛋白质组学研究及生物信息学分析[D].银川:宁夏医科大学,2013. LI J L.Study on proteomics and bioinformatics of the Echinococcus granulosus protoscoleces from infected humans[D].Yinchuan:Ningxia Medical University,2013.(in Chinese)
[32] DEA-AYUELA M A,ORDONEZ-GUTIERREZ L,BOLAS-FERNANDEZ F.Changes in the proteome and infectivity of Leishmania infantum induced by in vitro exposure to a nitric oxide donor[J].International Journal of Medical Microbiology,2009,299(3):221-232.
[33] ROBIJN M L,PLANKEN J,KORNELIS D,et al.Mass spectrometric detection of urinary oligosaccharides as markers of Schistosoma mansoni infection[J].Transactions of the Royal Society of Tropical Medicine and Hygiene,2008,102(1):79-83.
[34] KAWASE O,NISHIKAWA Y,BANNAI H,et al.Proteomic analysis off calcium-dependent secretion in Toxoplasma gondii[J].Proteomics,2007,7(20):3718-3725.
[35] CAO X,FU Z,ZHANG M,et al.iTRAQ-based comparative proteomic analysis of excretory-secretory proteins of schistosomula and adult worms of Schistosoma japonicum[J].Journal of Proteomics,2016,14(138):30-39.
[36] MA G,ZHANG J,YIN G,et al.Toxoplasma gondii:Proteomic analysis of antigenicity of soluble tachyzoite antigen[J].Experimental Parasitology,2009,122(1):41-46.
[37] ZHANG H,LEE E G,YU L,et al.Identification of the cross-reactive and species-specific antigens between Neospora caninum and Toxoplasma gondii tachyzoites by a proteomics approach[J].Parasitology Research,2011,109(3):899-911.
[38] BUDKE C M,DEPLAZES P,TORGERSON P R.Global socioeconomic impact of cystic echinococcosis[J].Emerging Infectious Diseases,2006,12(2):296-303.
[39] SMART O S,HORSKY V,GORE S,et al.Worldwide protein data bank validation information:Usage and trends[J].Acta Crystallographica Section D-Structural Biology,2018,74(3):237-244.
[40] TOWSE C L,AKKE M,DAGGETT V.The dynameomics entropy dictionary:A large-scale assessment of conformational entropy across protein fold space[J].The Journal of Physical Chemistry B,2017,121(16):3933-3945.
[41] PIEPER U,WEBB B M,DONG G Q,et al.ModBase,a database of annotated comparative protein structure models and associated resources[J].Nucleic Acids Research,2014,42:336-346.
[42] LOMIZE M A,POGOZHEVA I D,JOO H,et al.OPM database and PPM web server:Resources for positioning of proteins in membranes[J].Nucleic Acids Research,2012,40:370-376.
[43] ANDREEVA A,HOWORTH D,CHOTHIA C,et al.Investigating protein structure and evolution with SCOP2[J].Current Protocols in Bioinformatics,2015,49(1):1.26.1-1.26.21.
[44] WATERHOUSE A,BERTONI M,BIENERT S,et al.SWISS-MODEL:Homology modelling of protein structures and complexes[J].Nucleic Acids Research,2018,46(1):296-303.
[45] WARD M C,KOYFMAN S A.In reply to yildirim and topkan[J].International Journal of Radiation Oncology Biology Physics,2018,101(5):1273-1274.
[46] BOUTET E,LIEBERHERR D,TOGNOLLI M,et al.UniProtKB/Swiss-Prot,the manually annotated section of the UniProt knowledgeBase:How to use the entry view[J].Methods in Molecular Biology,2016,1374:23-54.
[47] APWEILER R,BAIROCH A,WU C H,et al.UniProt:The universal protein knowledgebase[J].Nucleic Acids Research,2004,32:115-119.
[48] CHEN B,PIEL W H,MONTEIRO A.Distal-less homeobox genes of insects and spiders:Genomic organization,function,regulation and evolution[J].Insect Science,2016,23(3):335-352.
[49] WOODS B,LANG B,BLAYNEY C,et al.Medic One Pediatric (MOPed) cards:Standardising paramedic paediatric resuscitation[J].BMJ Open Quality, 2019,8(3):534.
[50] FORREST A R,KAWAJI H,REHLI M,et al.A promoter-level mammalian expression atlas[J].Nature,2014,507(7493):462-470.
[51] FRENKEL-MORGENSTERN M,COHEN A A,GEVA-ZATORSKY N,et al.Dynamic Proteomics:A database for dynamics and localizations of endogenous fluorescently-tagged proteins in living human cells[J].Nucleic Acids Research,2010,38:508-512.
[52] MUTHUSAMY B,THOMAS J K,PRASAD T S,et al.Access guide to human proteinpedia[J].Current Protocols in Bioinformatics,2013,41(1):1.21.1-1.21.15.
[53] PEREZ-RIVEROL Y,CSORDAS A,BAI J,et al.The PRIDE database and related tools and resources in 2019:Improving support for quantification data[J].Nucleic Acids Research,2019,47(1):442-450.
[54] WANG M,HERRMANN C J,SIMONOVIC M,et al.Version 4.0 of PaxDb:Protein abundance data,integrated across model organisms,tissues,and cell-lines[J].Proteomics,2015,15(18):3163-3168.
[55] THUL P J,LINDSKOG C.The human protein atlas:A spatial map of the human proteome[J].Protein Science,2018,27(1):233-244.
[56] XENARIOS I,RICE D W,SALWINSKI L,et al.DIP:The database of interacting proteins[J].Nucleic Acids Research,2000,28(1):289-291.
[57] HAN K,PARK B,KIM H,et al.HPID:The human protein interaction database[J].Bioinformatics,2004,20(15):2466-2470.
[58] SIVADE D M,ALONSO-LOPEZ D,AMMARI M,et al.Encompassing new use cases-level 3.0 of the HUPO-PSI format for molecular interactions[J].BMC Bioinformatics, 2018,19(1):134-142.
[59] HOOGLAND C,MOSTAGUIR K,SANCHEZ J C,et al.SWISS-2DPAGE,ten years later[J].Proteomics,2004,4(8):2352-2356.
[60] OUGHTRED R,STARK C,BREITKREUTZ B J,et al.The BioGRID interaction database:2019 update[J].Nucleic Acids Research,2019,47(1):529-541.
[61] GILBERT D.Biomolecular interaction network database[J].Briefings in Bioinformatics,2005,6(2):194-198.
[62] KERRIEN S,ARANDA B,BREUZA L,et al.The IntAct molecular interaction database in 2012[J].Nucleic Acids Research,2012,40:841-846.
[63] LICATA L,BRIGANTI L,PELUSO D,et al.MINT,the molecular interaction database:2012 update[J].Nucleic Acids Research,2012,40:857-861.
[64] DAVIES M N,GUAN P,BLYTHE M J,et al.Using databases and data mining in vaccinology[J].Expert Opinion on Drug Discovery,2007,2(1):19-35.
[65] KAR P P,SRIVASTAVA A.Immuno-informatics analysis to identify novel vaccine candidates and design of a multi-epitope based vaccine candidate against theileria parasites[J].Frontiers in Immunology,2018,9:2213.
[66] 刘学磊,刘玉梅,曹文艳,等.细粒棘球绦虫AgB5抗原表位的生物信息学预测[J].中国病原生物学杂志,2015,10(1):38-41. LIU X L,LIU Y M,CAO W Y,et al.Bioinformatic prediction of the epitopes of Echinococcus granolusus antigen B5[J].Journal of Pathogen Biology,2015,10(1):38-41.(in Chinese)
[67] 张群,胡旭初,王卫国,等.日本血吸虫小蛋白型多药物/代谢物排出蛋白样基因(SMR-like)的发掘和生物信息学分析[J].热带医学杂志,2007,7(7):633-635,638. ZHANG Q,HU X C,WANG W G,et al.Uncovering and bioinformatics analysis of Schistosoma japonicum small multidrug/metabolites efflux protein-like gene[J].Journal of Tropical Medicine,2007,7(7):633-635.(in Chinese)
[68] SAPAY N,GUERMEUR Y,DELEAGE G.Prediction of amphipathic in-plane membrane anchors in monotopic proteins using a SVM classifier[J].BMC Bioinformatics, 2006,7:255.
[69] YACHDAV G,KLOPPMANN E,KAJAN L,et al.PredictProtein——An open resource for online prediction of protein structural and functional features[J].Nucleic Acids Research,2014,42:337-343.
[70] MOLLER S,CRONING M D,APWEILER R.Evaluation of methods for the prediction of membrane spanning regions[J].Bioinformatics,2001,17(7):646-653.
[71] BURLAND T G.DNASTAR’s Lasergene sequence analysis software[J].Methods in Molecular Biology,2000,132:71-91.
[72] SOLLNER J,GROHMANN R,RAPBERGER R,et al.Analysis and prediction of protective continuous B-cell epitopes on pathogen proteins[J].Immunome Research,2008,4:1.
[73] SAHA S,RAGHAVA G P.Prediction of continuous B-cell epitopes in an antigen using recurrent neural network[J].Proteins,2006,65(1):40-48.
[74] BEAVER J E,BOURNE P E,PONOMARENKO J V.EpitopeViewer:A java application for the visualization and analysis of immune epitopes in the immune epitope database and analysis resource (IEDB)[J].Immunome Research,2007,3:3.
[75] DICK T P,STEVANOVIC S,KEILHOLZ W,et al.The making of the dominant MHC classⅠ ligand SYFPEITHI[J].European Journal of Immunology,1998,28(8):2478-2486.
[76] PARKER K C,BEDNAREK M A,COLIGAN J E.Scheme for ranking potential HLA-A2 binding peptides based on independent binding of individual peptide side-chains[J].Journal of Immunology,1994,152(1):163-175.
[77] SIBLEY L D.Development of forward genetics in Toxoplasma gondii[J].International Journal for Parasitology,2009,39(8):915-924.
[78] WANG H L,ZHANG T E,YIN L T,et al.Partial protective effect of intranasal immunization with recombinant Toxoplasma gondii rhoptry protein 17 against toxoplasmosis in mice[J].PLoS One,2014,9(9):e108377.
[79] ROSA B,TOWNSEND R,JASMER D,et al.Functional and phylogenetic characterization of proteins detected in various nematode intestinal compartments[J].Molecular and Cellular Proteomics,2015,14(4):812-827.