Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (22): 4713-4719.doi: 10.3864/j.issn.0578-1752.2012.22.018

• VETERINARY SCIENCE • Previous Articles     Next Articles

Analysis of Immunological Characteristics of Echinococcus Granulosus Adult Worm Surface Membrane -Associated Glycoantigen

 GU  Nu-尔•Tu-尔Xun, MI  Xiao-Yun, ZHANG  Zhuang-Zhi, SHI  Bao-Xin, TU  尔Hong-•Yi-Mi-Ti, JIN  Ying-Hong, ZHANG  Mei, CHENG  Xiao-Bo, ZHANG  Xu, ZHAO  Li, ZHANG  Wen-Bao   

  1. 1.Veterinary Research Institute of Animal Science Academy of Xinjiang Uygur Autonomous Region，Urumqi 830000
    2.Xinjiang Agricultural University, Urumqi 830052
  • Received:2012-06-26 Online:2012-11-15 Published:2012-10-18

PDF

434

Abstract: 【Objective】 The immunological characteristics of the carbohydrate antigen (glycoprotein) of E. granulosus adult surface membrane were explored in this study . 【Method】 The freez-thaw E. granulosus adult worm lysis was digested by protease K, and centrifuged to collect supernatant as crude glycoantigen materials, which was identified by SDS-PAGE, quantified by sulfuric acid anthrone method. Then, the Balb/c mice were injected hypodermically by the material six times for anti-serum, which was collected to measure immunoglobulin (IgG, IgM and IgA) by ELISA, and to determine the specificity by western- blot method. To localize the site of the antigen composition in the E. granulosus adult worm by IFA. 【Result】 The SDS-PAGE showed that there was no visible band on the gel. This suggested that the proteins of the parasite surface membrane were degraded completely. The concentration of glycoantige was 2.54mg•mL-1 in the preparation. ELISA showed that the specific antibodies (IgG, IgM, and IgA) were rised gradually with the increase of the immunization times, and the positive/negative ration (P/N) of IgG, IgM and IgA equals to 4.9, 3.2 and 6.4, respectively. Comparatively, the specific IgE was not detected. Western blotting showed that the E. granulosus adult worm surface membrane glycoantigen and native proteins could be recognized by the anti-serum. IFA proved that the glycoantigen was located on the surface membrane of E. granulosus adult worm. 【Conclusion】 The glycoantigen of E. granulosus adult worms digested proteinase K has good immunogenicity and reactivity, and could be used to develop vaccine candidates and diagnosis material.

Key words: Echinococcus granulosus , surface membrane glycoantigen , antiserum

[1]张壮志, 石保新, 王进成, 吐尔洪•依米提, 哈斯也提, 哈江, 胡端铭, 李伯樵, 肉孜艾山, 吴 平, 王文明, 彭 政, 康强, 艾尔肯, 俞 进, 张文宝. 以家犬驱虫为中心的棘球蚴病控制措施在新疆两县的应用. 中国寄生虫与寄生虫病学杂志, 2008, 26(4): 253-257.

Zhang Z Z, Shi B X, Wang J Ch, Turhong Y, Hasiyet H, Hamalati J, Hu D M, Li B Q, Rouzi A, Wu P, Wang W M, Pen Z, Kang Q, Aikeng, Yu J, Zhang W B. Monthly deworming in dogs for Echinococcosis control in two counties of  Xinjiang Uygur Autonomous Region. Chinese Journal of Parasitology and Parasitic Disease, 2008, 26(4): 253-257. (in Chinese)

[2]张壮志, 张文宝, 石保新, 王进成, 王力俭. 我国包虫病防控及其面临的困难. 兽医导刊, 2011, 6: 27-29.

Zhang Z Z, Zhang W B, Shi B X, Wang J C, Wang L J. Hydatid disease prevention and control in China and its difficulties. Veterinary Orientation, 2011, 6: 27-29. (in Chinese)

[3]Cecilia C, Ramiro M, Carlos C. Characterization of carbohydrates of adult Echinococcus granulosus by lectin-binding analysis. The Joural of Parasitology, 2003, 89(1): 57-61.

[4]Desiree V D K, Alexandra V R, Joost H. N S, Martien L K, Andre M D, Aloysius G. M. T, Cornelus H. H, Maria Y. Triggering of innate immune responses by schistosome egg glycolipids and their carbohydrate epitope GalNAcb1-4(Fuca1-2Fuca1-3)GlcNAc. The Journal of Infectious Diseases, 2002, 185: 531-539.

[5]Andrew J R, Lauri A E, Judith A A, Nancy W, Robert B G, Michael M, Donald L W, Howard R M, Anne D. Novel tyvelose-containing tri- and tetra-antennary N-glycans in the immunodominant antigens of the intracellular parasite trichinella spiralis. Glycobiology, 1994, 4: 593-603.

[6]Marina. M, Adriana B, Alberto N. Carbohydrates on the surface of Echinococcus granulosus protoescoleces are immunodominant in mice. Parasite Immunology, 1996, 18(11): 559-569.

[7]Nyame A K, Kawar Z S, Richard D C. Antigenic glycans in parasitic infections: implications for vaccines and diagnostics. Archives of Biochemistry and Biophysics, 2004: 426, 182-200.

[8]张 英, 黄林娟, 王仲孚. 糖蛋白的凝胶电泳和电印迹染色鉴定技术. 化学进展, 2008, 20(7/8): 1158-1164.

Zhang Y, Huang L J, Wang Z F. Techniques for staining glycoprotein on Gel electrophoresis or electroblotting. Progress In Chemistry, 2008, 20(7/8): 1158-1164. (in Chinese)

[9]Katrin H, Joachim K. Surface carbohydrate composition of a tapeworm in its consecutive intermediate hosts: Individual variation and fitness consequences. International Journal for Parasitology, 2005, 35: 1499-1507.

[10]古努尔. 吐尔逊, 米晓云, 张壮志, 雷震, 石保新, 巫 剑, 赵 莉, 易 忠, 吐尔洪•依米提, 张文宝. 细粒棘球绦虫成虫表膜抗原特异性基因的筛选及克隆. 中国动物传染病学报, 2009, 17(4): 47-51.

Gulnur T, Mi X Y, Zhang Z Z, Lei Z, Shi B X, Yi Zh, Turhong •Y, Zhang W B. Immunoscreening and cloning of tegumental surface antigen of Echinococcus granulosus adult worm. Chinese Journal of Animal Infectious Diseases, 2009, 17(4): 47-51. (in Chinese)

[11]古努尔•吐尔逊, 米晓云, 张壮志, 石保新, 吐尔洪•依米提, 张 旭, 巫 剑, 赵 莉, 阿曼古丽•马木提, 金映红, 张文宝. 用抗虫体表膜抗原抗体测定犬细粒棘球绦虫粪抗原. 畜牧兽医学报, 2011, 42(6): 845-850.

Gulnur T, Mi X Y, Zhang Z Z, Shi B X, Turhong Y, Zhang X, Wu J, Zhao L, Amangul M, Jing Y H, Zhang W B. Using antibodies against tegument antigens for Copro-ELISA to detect Echinococcus granulosus in dogs. Acta Veterinaria et Zootechnica Sinica, 2011, 42(6): 845-850. (in Chinese)

[12]黄 燕, Heath D David, Antone J Christine, 邱加闽, 王 谦, 陈兴旺, 黄 亮, 易德友, 杨 文, 杨 筠. 犬粪便棘球绦虫抗原的双抗体夹心酶联免疫吸附试验检测法建立与应用. 预防医学情报杂志, 2008, 24(6): 407-411.

Huang Y, Heath D D, Antone J C, Qiu J M, Wang Q, Cheng X W, Huang L, Yi D Y, Yang W, Yang Y. An immunological coproantigen test used to detect infection of Echinococcus in dogs. Journal of Preventive Medicine information, 2008, 24(6): 407-411. (in Chinese)

[13]Huang Y, Yang W, Qiu J M, Chen X W, Yang Y, Qiu D C, Xiao N, Xiao Y F, David H. A modified coproantigen test used for surveillance of Echinococcus spp. in Tibetan dogs. Veterinary Parasitology, 2007, 149: 229-238.

[14]Elayoubi F A, Fraser A, Jenkins D J, Craig P S. Partial characterisation of carbohydrate-rich Echinococcus granulosus antigens. International Journal for Parasitology, 2003, 33: 1553-1559.

[15]Elayoubi F A, Craig P S. Echinococcus granulosus coproantigens: chromatographic fractionation and characterisation. Parasitology, 2004, 128: 455-465.

[16]Jacobson R L, Doyle R J. Lectin parasite interactions. Parasitology Today, 1996, 12: 55-61.

[17]Gasque P. Complement: a unique innate immune sensor for danger signals. Molecular Immunology, 2004, 41: 1089-1098.

[18]Alvaro D, Fontana E C, Todeschini A R. The major surface carbohydrates of the Echinococcus granulosus cyst: Mucin-Type O-Glycans decorated by novel galactose-based structures. Biochemistry, 2009, 48, 11678-11691.

[19]石保新, 巫 剑, 张壮志, 张文宝, 米晓云, 古努尔. 吐尔逊, 岳城, 吐尔洪. 依米提, 张 旭, 赵 莉, 郝呈普. EgM9蛋白免疫犬肠系膜淋巴结中抗体的免疫荧光组织化学定位. 中国兽医科学, 2010, 40(04): 406-409.

Shi B X, Wu J, Zhang Z Z, Zhang W B, Mi X Y, Gulnur. T, Yue C, Torhong. Y, Zhang X, Zhao L, Hao C P. Localization of specific antibody inmesenteric lymph nodes of dogs vaccinated with EgM9 protein from Echinococus granulosus by immunofluorescence technique. Chinese Veterinary Science, 2010, 40(04): 406-409. (in Chinese)

[20]Ari H, Markus A. Intracellular functions of N-linked glycans. Science, 2001, 291(5512): 2364-2369.

[21]Ohtsubo K, Marth J D. Glycosylation in cellular mechanisms of mechanisms of health and disease. Cell, 2006, 126(5): 855-867.

[22]卢 庄, 王京兰, 蔡 耘, 钱小红. 糖蛋白组学研究技术进展及其应用. 生命的化学, 2008, 28(3): 335-338.

Lu Z, Wang J L, Cai  Y, Qian X H. Progress of research on glycoprotein and its application. Chemistry of Life, 2008, 28(3): 335-338. (in Chinese)

[23]沈锦玉, 余旭平, Kim Thompson, Alexandra Adams. 嗜水气单胞菌细胞提取物特性的分析. 浙江大学学报（农业与生命学报）, 2005, 31(5): 555-560.

Shen J Y, Yu X P, Kim T, Alexandra A. Characterization of extract of aeromonas hydrophila. Journal of Zhejiang University(Agric & Life Sci. ), 2005, 31(5): 555-560. (in Chinese)

[24]张丽芳, 刘　星, 李　红. 嗜肺巴氏杆菌外膜蛋白和脂多糖提取及其活性的初步研究. 中国比较医学杂志, 2003, 13(5): 300-304.

Zhang L F, Liu X, Li  H. Preliminary study on the extraction and activity of outer membrane protein and lipopolysaccharide of pasteuralla pneumotropica. Chinese Journal of Comparative Medicine, 2003, 13(5): 300-304. (in Chinese)

[25]张 倩, 杨 振, 张艳贞, 王爱丽, 安学丽, 晏月明. 蛋白质糖基化修饰的研究方法及其应用. 生物技术通报, 2006, 1: 46-49.

Zhang Q, Yang Z, Zhang Y Z, Wang A L, Yan X L, Yan Y M. Detection of protein glycosylation modifications and its applications. Biotechnology Bulletin, 2006, 1: 46-49. (in Chinese)　

[26]高雅楠, 田 芳, 陈 琳, 季旻珺, 吴观陵. 日本血吸虫糖基诱导抗体类型与感染状态的关系研究. 中国血吸虫病防治杂志, 2010, 22(6): 528-533.

Gao Y N, Tian F, Chen L, Ji M J, Wu G L. Study on role of carbohydrate-specific antibodies in pigs exposed to normal and/or attenuated Schistosoma japonicum cercariae. Chinese Journal of  Schistosomiasis Control, 2010, 22(6): 528-533. (in Chinese)

[27]Cynthia W A N, AlexandraV R, John H. O, Gachuhi K, David W D, Johannis P K, Andre M D, Cornelis H H. Specific antibody responses to three schistosome-related carbohydrate strctures in recently exposed immigrants and established residents in an area of Schistosoma mansoni endemicity. Infection and Immunity, 2003, 71(10): 5676-5681.

[28]Alexandra V R, Govert J V D, Cornelis H H, Dirk H V D E, Iram V D, Andre M D. Profiles of immunoglobulin M(IgM) and IgG antibodies against defined carbohydrate epitopes in sera of Schistosoma-infected individuals determined by surface Plasmon reasonance. Infection and Immunity, 2001, 69(4): 2396-2401.
[1] JIA Guo-Chao-1, 2 , ZHI Ai-Min-1, LI Meng-Qin-2, SONG Chun-Mei-1, WANG Ling-Ling-1, LIU Ru-Biao-1, HU Xiao-Fei-1, WANG Fang-Yu-1, ZHANG Gai-Ping-1. Study on Rapid Detection Kit of Fleroxacin by icELISA [J]. Scientia Agricultura Sinica, 2014, 47(11): 2251-2261.
[2] ZHAO Li, CHEN Hao-Fei, ZHANG Wen-Bao, MA Zheng-Hai, ZHANG Zhuang-Zhi, ZHANG Xu, GU Nu-尔?Tu-尔Xun, MI Xiao-Yun, JIN Ying-Hong, XUE Jing, SHI Bao-Xin. Cloning, Prokaryotic Expression of Echinococcus granulosus Heat Shock Protein 70 and Preparation of It’s Antiserum [J]. Scientia Agricultura Sinica, 2012, 45(12): 2491-2501.
[3] HU Xiao-fei,WEI Feng-xian,XING Guang-xu,ZHI Ai-min,CHAI Shu-jun,SUN Ya-ning,WANG Lei,DENG Rui-guang,ZHANG Gai-ping
. Preparation of Phytase Mouse Polyclonal Antiserum
 [J]. Scientia Agricultura Sinica, 2011, 44(3): 613-619 .
[4] JIA Yong-jian,FENG Bao-zhen,LI Pei-qian,ZHANG Xiu-guo,FU Hong-bo
. Eukaryotic Expression and Functional Analysis of Pcpme3 From Phytophthora capsici
 [J]. Scientia Agricultura Sinica, 2009, 42(6): 1988-1993 .
[5] . Development of antibodies against Diethylstilbestrol [J]. Scientia Agricultura Sinica, 2008, 41(5): 1524-1529 .
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd