Prediction and yeast surface display of antigenic epitope region of TgMIC16 of Toxoplasma gondii
SUN Hui1, LI Jin1, XIAO Ting1, XU Chao1, YIN Kun1, HUANG Bing-cheng1, LEI Zhan2, WEI Qing-kuan1
1. Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Diseases, Jining 272033, China; 2. Shandong Academy of Medical Sciences, Shandong Laboratory Animal Center, Jinan 250000, China
Abstract:The bioinformatics software was used to predict the B cell and T cell epitopes of Toxoplasma gondii microneme 16 (TgMIC16) followed by epitopes display on the yeast of Saccharomyces cerevisiae. B and T cell epitopes of TgMIC16 were predicted by DNAStar and IEDB, and software of SYFPEITHI and BIMAS, respectively. According to the predicted results, the antigenic epitope region was expressed with pCTCON2/EBY100 display system. The expression protein was detected by indirect immunofluorescence (IFA) and flow cytometry. Results showed that there were potential B/T cell epitopes in TgMIC16 and concentrated in the aa343-625 region. The epitope region was successfully displayed on the surface of yeast cells, and the optimal induction time was 24 hours. The study would lay the foundation for the development and efficacy evaluation of the yeast carrier vaccine in the further research.
基金资助:国家自然科学基金(No.81501770); 山东省自然科学基金(No.ZR2015YL051, No.ZR2015YL033, No ZR2016CP18); 山东省医学科学院医药卫生科技创新工程
通讯作者:
魏庆宽,Email: wqkzlj@sina.com
引用本文:
孙慧, 李瑾, 肖婷, 徐超, 尹昆, 黄炳成, 雷战, 魏庆宽. 弓形虫微线蛋白16抗原表位区的预测及酵母表面展示[J]. 中国人兽共患病学报, 2017, 33(9): 774-778.
SUN Hui, LI Jin, XIAO Ting, XU Chao, YIN Kun, HUANG Bing-cheng, LEI Zhan, WEI Qing-kuan. Prediction and yeast surface display of antigenic epitope region of TgMIC16 of Toxoplasma gondii. Chinese Journal of Zoonoses, 2017, 33(9): 774-778.
[1] Saouros S, Dou Z, Henry M, et al. Microneme protein 5 regulates the activity of Toxoplasma subtilisin 1 by mimicking a subtilisin prodomain[J]. J Biol Chem, 2012, 287(43): 36029-36040. doi:10.1074/jbc.M112.389825 [2] Wang T, Tang ZH. Toxoplasmosis: tHe spread of tachyzoites, cysts and oocysts[J]. Chin J Zoonoses, 2012, 28(11): 1133-1136. (in Chinese) 汪涛,汤自豪. 弓形虫病:速殖子、包囊和卵囊的传播[J]. 中国人兽共患病学报, 2012, (11):1133-1136. [3] Torrey EF, Bartko JJ, Yolken RH. Toxoplasma gondii and other risk factors for schizophrenia: an update[J]. Schizophr Bull, 2012, 38(3): 642-647. doi:10.1093/schbul/sbs043 [4] Hoff EF, Cook SH, Sherman GD, et al. Toxoplasma gondii : molecular cloning and characterization of a novel 18-kDa secretory antigen, TgMIC10[J]. Exp Parasitol, 2001, 97(2): 77-88. doi:10.1006/expr.2000.4585 [5] Lourenco EV, Bernardes ES, Silva NM, et al. Immunization with MIC1 and MIC4 induces protective immunity against Toxoplasma gondii [J]. Microbes Infect, 2006, 8: 1244-1251. doi:10.1016/j.micinf.2005.11.013 [6] Fang R, Nie H, Wang ZS, et al. Protective immune response in BALB/c mice induced by a suicidal DNA vaccine of the MIC3 gene of Toxoplasma gondii [J]. Vet Parasitol, 2009, 164: 134-140. doi:10.1016/j.vetpar.2009.06.012 [7] Peng GH, Yuan ZG, Zhou DH, et al. Toxoplasma gondii microneme protein 6 (MIC6) is a potential vaccine candidate against toxoplasmosis in mice[J]. Vaccine, 2009, 27: 6570-6574. doi:10.1016/j.vaccine.2009.08.043 [8] Liu MM, Yuan ZG, Peng GH, et al. Toxoplasma gondii microneme protein 8 (MIC8) is a potential vaccine candidate against toxoplasmosis[J]. Parasitol Res, 2010, 106: 1079-1084. doi:10.1007/s00436-010-1742-0 [9] Shin MK, Yoo HS. Animal vaccines based on orally presented yeast recombinants[J]. Vaccine, 2013, 31(40): 4287-4292. doi:10.1016/j.vaccine.2013.07.029 [10] Li J, Cui Y, Yin K, et al. Prokaryotic expression, purification and immunological characterization of micronemal protein 16 of toxoplama gondii [J]. Chin J Parasitol Parasit Dis, 2016, 34(3): 198-202. (in Chinese) 李瑾,崔勇,尹昆,等. 刚地弓形虫微线蛋白16基因片段原核表达、纯化及免疫反应性分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(3):198-202. [11] Li YJ, Wang J, Zhao H, et al. Bioinformatics prediction on eg95 antigen epitopes of Echinococcus granulosus [J]. Chin J Zoonoses, 2011, 27(10): 892-896, 900. (in Chinese) 李玉娇,王晶,赵慧,等. 细粒棘球绦虫 Eg 95抗原表位的生物信息学预测[J]. 中国人兽共患病学报,2011,(10):892-896,900. [12] He SW, Wei XX. Prediction of epitopes of the myophilin protein from Echinococcus granulosus [J]. J Pathog Biol, 2017, 12(1): 42-45,50. (in Chinese) 何顺伟,魏晓星. 细粒棘球绦虫myophilin的抗原表位预测[J]. 中国病原生物学杂志,2017,(01):42-45,50. [13] Lee SY, Choi JH, Xu Z. Microbial cell-surface display[J]. Trends Biotechnol, 2003, 21(1): 45-52. doi:10.1016/S0167-7799(02)00006-9 [14] Boder ET, Wittrup KD. Yeast surface display for screening combinatorial polypeptide libraries[J]. Nat Biotechnol, 1997, 15: 553-557. doi:10.1038/nbt0697-553