Immune response in guinea pigs vaccinated with DNA vaccine of foot-and-mouth disease virus O/China99
Immune response in guinea pigs vaccinated with DNA vaccine of foot-and-mouth disease virus O/China99
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DOI：
10.1016/j.vaccine.2004.03.074
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发表时间：
2005-05-09
期刊：
影响因子：
5.5
通讯作者：
Xie, QG
中科院分区：
文献类型：
--
作者：
Guo, HC;Liu, ZX;Xie, QG
为了获得包含全长P1、2A、3C和部分2B、3B的口蹄疫病毒(FMDV O/China99)基因P12X3C，采用位点突变策略。将重组质粒pcDNA3.1/P12X3C转染BHK-21细胞。通过夹心ELISA和间接免疫荧光试验证实了BHK-21细胞中表达的FMDV衣壳蛋白。然后将质粒pcDNA3.1/P12X3C肌肉注射给豚鼠，并将酵母细胞中表达的纯化FMDV O/China993D蛋白与pcDNA3.1/P12X3C一起注射。采用间接ELISA法检测抗FMDV抗体，采用MTT法检测T淋巴细胞增殖反应，采用微量中和法分析中和抗体滴度。结果表明，质粒pcDNA3.1/P12X3C能够在BHK-21细胞中表达FMDV免疫活性蛋白。此外，在疫苗接种后第二周，质粒pcDNA3.1/P12X3C引发并增加了抗FMDV抗体。接种疫苗后可诱导中和抗体，增强 T 淋巴细胞增殖反应。在攻毒试验中，所有接种pcDNA3.1/P12X3C的豚鼠均得到充分保护，免受FMDV攻毒。然而，从注射蛋白质3D和质粒pcDNA3.1/P12X3C的动物获得的结果并不令人满意。总之，该结果鼓励进一步开展口蹄疫DNA疫苗的开发工作，并为DNA疫苗的研究奠定基础。 (c) 2004 Elsevier Ltd. 保留所有权利。
In order to obtain the gene P12X3C of foot-and-mouth disease virus (FMDV O/China99) that includes full length P1, 2A, 3C and part of 2B and 3B, the site mutation strategy was used. The recombinant plasmid pcDNA3.1/P12X3C was transfected into BHK-21 cells. The capsid proteins of FMDV expressed in BHK-21 cells were confirmed by sandwich-ELISA and indirect immunofluorescence test. Then the plasmid pcDNA3.1/P12X3C was administered to guinea pigs intramuscularly, and purified FMDV O/China993D protein expressed in yeast cells was injected together with pcDNA3.1/P12X3C. Anti-FMDV antibodies were detected by indirect ELISA, the T-lymphocyte proliferation response was tested by MTT assay, and neutralizating antibodies titers were analyzed by micro-neutralization assay. The result showed that the plasmid pcDNA3.1/P12X3C was able to express immunocompetent proteins of FMDV in BHK-21 cells. Furthermore, anti-FMDV antibodies were elicited and increased by plasmid pcDNA3.1/P12X3C in the second week after vaccination. Neutralizating antibodies were induced and the T-lymphocyte proliferation response was enhanced after vaccination. In the challenge test, all of guinea pigs vaccinated with pcDNA3.1/P12X3C were fully protected from FMDV challenge. However, the result obtained from animals that were injected with protein 3D together with plasmid pcDNA3.1/P12X3C was not satisfied. In conclusion, the results encouraged further work towards the development of a DNA vaccine against FMDV and provided the basis of research for DNA vaccine. (c) 2004 Elsevier Ltd. All rights reserved.