背景:绵羊肺腺癌(OPA)是由Jaagsiekte逆转录病毒(JSRV)引起的绵羊传染性肺癌。OPA通常在世界范围内具有严重的经济影响。疫苗尚未开发出来,尽管这种疾病已经在全球传播,连同它的并发症。本研究旨在利用免疫信息学工具构建针对JSRV诱导B和T淋巴细胞的有效多表位疫苗。
结果:设计的疫苗由499个氨基酸组成。在疫苗被计算验证之前,考虑了所有关键参数;包括抗原性,变应原性,毒性,和稳定性。疫苗的理化性质显示出9.88的等电点。根据不稳定指数(II),疫苗稳定在28.28。该疫苗在脂肪族指数上得分为56.51,在GRAVY上得分为-0.731,表明疫苗是亲水的。RaptorX服务器用于预测疫苗的三级结构,GalaxyWEB服务器改进了结构,Ramachandran图和ProSA-Web服务器验证了疫苗的三级结构。蛋白质溶胶和SOLPro服务器显示了疫苗的溶解度。此外,二硫化物工程减少了疫苗结构中的高流动性区域,提高了疫苗的稳定性。此外,疫苗构建体与绵羊MHC-1等位基因对接,并显示出有效的结合能.免疫模拟显着显示高水平的免疫球蛋白,T淋巴细胞,和INF-γ分泌物。分子动力学模拟提供了构建疫苗的稳定性。最后,将疫苗反转录成DNA序列,并克隆到pET-30a(+)载体中,以确认翻译效力和微生物表达.
结论:一种针对JSRV的新型多表位疫苗构建体,由B和T淋巴细胞表位形成,并产生了潜在的保护。这项研究可能有助于控制和根除OPA。
BACKGROUND: Sheep pulmonary adenocarcinoma (
OPA) is a contagious lung cancer of sheep caused by the Jaagsiekte retrovirus (JSRV).
OPA typically has a serious economic impact worldwide. A vaccine has yet to be developed, even though the disease has been globally spread, along with its complications. This study aimed to construct an effective multi-epitopes vaccine against JSRV eliciting B and T lymphocytes using immunoinformatics tools.
RESULTS: The designed vaccine was composed of 499 amino acids. Before the vaccine was computationally validated, all critical parameters were taken into consideration; including antigenicity, allergenicity, toxicity, and stability. The physiochemical properties of the vaccine displayed an isoelectric point of 9.88. According to the Instability Index (II), the vaccine was stable at 28.28. The vaccine scored 56.51 on the aliphatic index and -0.731 on the GRAVY, indicating that the vaccine was hydrophilic. The RaptorX server was used to predict the vaccine\'s tertiary structure, the GalaxyWEB server refined the structure, and the Ramachandran plot and the ProSA-web server validated the vaccine\'s tertiary structure. Protein-sol and the SOLPro servers showed the solubility of the vaccine. Moreover, the high mobile regions in the vaccine\'s structure were reduced and the vaccine\'s stability was improved by disulfide engineering. Also, the vaccine construct was docked with an ovine MHC-1 allele and showed efficient binding energy. Immune simulation remarkably showed high levels of immunoglobulins, T lymphocytes, and INF-γ secretions. The molecular dynamic simulation provided the stability of the constructed vaccine. Finally, the vaccine was back-transcribed into a DNA sequence and cloned into a pET-30a ( +) vector to affirm the potency of translation and microbial expression.
CONCLUSIONS: A novel multi-epitopes vaccine construct against JSRV, was formed from B and T lymphocytes epitopes, and was produced with potential protection. This study might help in controlling and eradicating
OPA.