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Design and production of dengue virus chimeric proteins useful for developing tetravalent vaccines.
Vaccine ( IF 5.5 ) Pub Date : 2020-01-22 , DOI: 10.1016/j.vaccine.2020.01.003 Izabella Cristina Andrade Batista 1 , Bárbara Resende Quinan 2 , Érica Alessandra Rocha Alves 1 , Soraya Torres Gaze Jangola 1 , Eneida Santos Oliveira 1 , Stella Garcia Colombarolli 1 , Jorge Gomes Goulart Ferreira 1 , Eliseu Soares de Oliveira Rocha 3 , Erna Geessien Kroon 4 , Rafael Ramiro de Assis 5 , Jaquelline Germano de Oliveira 1 , Jacqueline Araújo Fiuza 1 , Carlos Eduardo Calzavara-Silva 1
Vaccine ( IF 5.5 ) Pub Date : 2020-01-22 , DOI: 10.1016/j.vaccine.2020.01.003 Izabella Cristina Andrade Batista 1 , Bárbara Resende Quinan 2 , Érica Alessandra Rocha Alves 1 , Soraya Torres Gaze Jangola 1 , Eneida Santos Oliveira 1 , Stella Garcia Colombarolli 1 , Jorge Gomes Goulart Ferreira 1 , Eliseu Soares de Oliveira Rocha 3 , Erna Geessien Kroon 4 , Rafael Ramiro de Assis 5 , Jaquelline Germano de Oliveira 1 , Jacqueline Araújo Fiuza 1 , Carlos Eduardo Calzavara-Silva 1
Affiliation
Dengue virus (DENV) is a Flavivirus estimated to cause 390 million infections/year. Currently, there is no anti-viral specific treatment for dengue, and efficient DENV vector control is still unfeasible. Here, we designed and produced chimeric proteins containing potential immunogenic epitopes from the four DENV serotypes in an attempt to further compose safer, balanced tetravalent dengue vaccines. For this, South American DENV isolate sequences were downloaded from the NCBI/Virus Variation/Dengue virus databases and intraserotype-aligned to generate four consensuses. Four homologous DENV sequences were retrieved using BLAST and then interserotype-aligned. In parallel, sequences were subjected to linear B epitope prediction analysis. Regions of the envelope and NS1 proteins that are highly homologous among the four DENV serotypes, non-conserved antigenic regions and the most antigenic epitopes found in the C, prM, E and NS1 DENV proteins were used to construct 11 chimeric peptides. Genes encoding the chimeric proteins were commercially synthesized, and proteins were expressed, purified by affinity chromatography and further subjected to ELISA assays using sera from individuals infected with DENVs 1, 2, 3 or 4. As a proof-of-concept, the chimeric EnvEpII protein was selected to immunize BALB/c and C57BL/6 mice strains. The immunization with EnvEpII protein associated with aluminum induced an increased number of T CD4+ and CD8+ cells, high production of IgG1 and IgG2 antibodies, and increased levels of IL-2 and IL-17 cytokines, in both mouse strains. Because the EnvEpII protein associated with aluminum induced an efficient cellular response by stimulating the production of IL-2, IL-4, IL-17 and induced a robust humoral response in mice, we conclude that it resembles an efficient specific response against DENV infection. Although further experiments are required, our results indicate that epitope selection by bioinformatic tools is efficient to create recombinant proteins that can be used as candidates for the development of vaccines against infectious diseases.
中文翻译:
设计和生产可用于开发四价疫苗的登革热病毒嵌合蛋白。
登革热病毒(DENV)是一种黄病毒,估计每年引起3.9亿次感染。当前,没有针对登革热的抗病毒特异性治疗,并且仍然不能有效地控制登革热载体。在这里,我们设计并生产了包含来自四种DENV血清型的潜在免疫原性表位的嵌合蛋白,以试图进一步组成更安全,平衡的四价登革热疫苗。为此,从NCBI /病毒变异/登革热病毒数据库下载了南美DENV分离株序列,并进行了血清内比对,以产生四个共识。使用BLAST检索了四个同源DENV序列,然后进行了血清型比对。并行地,对序列进行线性B表位预测分析。四种DENV血清型之间高度同源的包膜和NS1蛋白区域,在C,prM,E和NS1 DENV蛋白中发现的非保守抗原区和最具抗原性的抗原决定簇被用于构建11个嵌合肽。商业合成编码嵌合蛋白的基因,表达蛋白,通过亲和层析纯化,并进一步使用来自被DENV 1、2、3或4感染的个体的血清进行ELISA测定。作为概念验证,嵌合EnvEpII选择蛋白以免疫BALB / c和C57BL / 6小鼠品系。在两种小鼠品系中,与铝相关的EnvEpII蛋白免疫诱导了T CD4 +和CD8 +细胞数量的增加,IgG1和IgG2抗体的高产量以及IL-2和IL-17细胞因子的水平增加。因为与铝有关的EnvEpII蛋白通过刺激IL-2,IL-4,IL-17的产生诱导了有效的细胞反应,并在小鼠中诱导了强烈的体液反应,所以我们得出结论,它类似于针对DENV感染的有效特异性反应。尽管需要进行进一步的实验,但我们的结果表明,通过生物信息学工具对抗原决定簇的选择有效地产生了重组蛋白,可以用作开发抗传染病疫苗的候选蛋白。
更新日期:2020-01-23
中文翻译:
设计和生产可用于开发四价疫苗的登革热病毒嵌合蛋白。
登革热病毒(DENV)是一种黄病毒,估计每年引起3.9亿次感染。当前,没有针对登革热的抗病毒特异性治疗,并且仍然不能有效地控制登革热载体。在这里,我们设计并生产了包含来自四种DENV血清型的潜在免疫原性表位的嵌合蛋白,以试图进一步组成更安全,平衡的四价登革热疫苗。为此,从NCBI /病毒变异/登革热病毒数据库下载了南美DENV分离株序列,并进行了血清内比对,以产生四个共识。使用BLAST检索了四个同源DENV序列,然后进行了血清型比对。并行地,对序列进行线性B表位预测分析。四种DENV血清型之间高度同源的包膜和NS1蛋白区域,在C,prM,E和NS1 DENV蛋白中发现的非保守抗原区和最具抗原性的抗原决定簇被用于构建11个嵌合肽。商业合成编码嵌合蛋白的基因,表达蛋白,通过亲和层析纯化,并进一步使用来自被DENV 1、2、3或4感染的个体的血清进行ELISA测定。作为概念验证,嵌合EnvEpII选择蛋白以免疫BALB / c和C57BL / 6小鼠品系。在两种小鼠品系中,与铝相关的EnvEpII蛋白免疫诱导了T CD4 +和CD8 +细胞数量的增加,IgG1和IgG2抗体的高产量以及IL-2和IL-17细胞因子的水平增加。因为与铝有关的EnvEpII蛋白通过刺激IL-2,IL-4,IL-17的产生诱导了有效的细胞反应,并在小鼠中诱导了强烈的体液反应,所以我们得出结论,它类似于针对DENV感染的有效特异性反应。尽管需要进行进一步的实验,但我们的结果表明,通过生物信息学工具对抗原决定簇的选择有效地产生了重组蛋白,可以用作开发抗传染病疫苗的候选蛋白。