背景:A型流感病毒具有传染性,如果不及时治疗,甚至会危及生命。到目前为止,由于快速的抗原变化和新亚型流感病毒的出现,目前尚无广泛的保护性疫苗.在这项研究中,我们利用生物信息学工具,从H7N9亚型流感病毒HA和M2蛋白的抗原区和高度保守区设计亚单位嵌合疫苗.我们使用了粘膜佐剂候选物,包括CTxB,STxB,ASP-1和LTB刺激粘膜免疫,并分析了HA2,M2e,和佐剂。此外,为了改善抗原功能并保持其三维结构,使用12种不同的接头,包括6种刚性接头和6种柔性接头。采用同源和从头算相结合的建模方法生成三维结构模型,并对模型的分子动力学进行分析,要么。
结果:对不同佐剂的分析表明,使用CtxtB作为佐剂,在4种候选佐剂中,疫苗的总体稳定性和半衰期均较高.与其他组合形式相比,抗原和M2e-接头-CTxB-接头-HA2形式的CTxB的融合具有最大的稳定性和半衰期。此外,在12个分析的接头中,KPKPKP刚性接头显示该候选疫苗的最佳结果.通过连接体插入疫苗3D结构的变化可以忽略不计,然而,虽然小,抗原之间的接头插入导致结构轻微变化。最终,使用诸如Ellipro之类的预测工具,NetMHCpanI和II,CD4episcore,CTLpred,BiepPred和其他表位分析工具,我们分析了疫苗的构象和线性表位。溶解度,蛋白酶体切割位点,肽酶和潜在的化学切割器,密码子优化,翻译后修饰也在最终的疫苗上进行。
结论:结论:当使用KPKPKP作为接头和使用CTxB作为佐剂时,M2e-接头-CTxB-接头-HA2组合的嵌合疫苗保留了其3D结构和抗原性。
BACKGROUND: Type A influenza viruses are contagious and even life-threatening if left untreated. So far, no broadly protective vaccine is available due to rapid antigenic changes and emergence of new subtypes of influenza virus. In this
study, we exploited bioinformatics tools in order to design a subunit chimeric vaccine from the antigenic and highly conserved regions of HA and M2 proteins of H7N9 subtype of influenza virus. We used mucosal adjuvant candidates, including CTxB, STxB, ASP-1, and LTB to stimulate mucosal immunity and analyzed the combination of HA2, M2e, and the adjuvant. Furthermore, to improve the antigen function and to maintain their three-dimensional structure, 12 different linkers including six rigid linkers and six flexible linkers were used. The 3D structure model was generated using a combination of homology and ab initio modeling methods and the molecular dynamics of the model were analyzed, either.
RESULTS: Analysis of different adjuvants showed that using CtxB as an adjuvant, results in higher overall vaccine stability and higher half-life among four adjuvant candidates. Fusion of antigens and the CTxB in the form of M2e-linker-CTxB-linker-HA2 has the most stability and half life compared to other combination forms. Furthermore, the KPKPKP rigid linker showed the best result for this candidate vaccine among 12 analyzed linkers. The changes in the vaccine 3D structure made by linker insertion found to be negligible, however, although small, the linker insertion between the antigens causes the structure to change slightly. Eventually, using predictive tools such as Ellipro, NetMHCpan I and II, CD4episcore, CTLpred, BepiPred and other epitope analyzing tools, we analyzed the conformational and linear epitopes of the vaccine. The solubility, proteasome cleavage sites, peptidase and potential chemical cutters, codon optimization, post translational modification were also carried out on the final vaccine.
CONCLUSIONS: It is concluded that M2e-Linker-CTxB-Linker-HA2 combination of chimeric vaccine retains its 3D structure and antigenicity when KPKPKP used as linker and CTxB used as adjuvant.