Department of Pharmaceutical Biomaterials, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran; Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
Department of Pharmaceutical Biomaterials, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
Int Immunopharmacol. 2021 Feb;91:107265. doi: 10.1016/j.intimp.2020.107265. Epub 2020 Dec 25.
Glioblastoma multiform is the most common of primary malignant brain tumors in adults. Currently, surgical resection of the tumor mass, followed by adjuvant radiotherapy and chemotherapy are standard treatments for glioblastoma multiform but so far are not effective treatments. Thus, the development of a vaccine, as a safe and efficient strategy for prophylactic or therapeutic purposes against glioblastoma multiform is very necessary. The present study aimed to design the multi-domain vaccine for glioblastoma multiform. An in silico approach was used to select the most potent domains of proteins to induce the host's B- and T-cell immune response against glioblastoma multiform. IL-13Rα-2 (amino acid positions 27-144), TNC (amino acid positions 1900-2100), and PTPRZ-1(amino acid positions 731-884) were found to have potent inducible immune responses. So, we considered them for fusing with a linker A(EAAAK)A to construct the multi-domain recombinant vaccine. The immuno-informatics analysis of the designed recombinant vaccine construct was performed to evaluate its efficacy. Although the designed recombinant vaccine construct did not show allergen property, its antigenicity was estimated at 0.78. The Physico-chemical properties of the recombinant vaccine construct were characterized and revealed the potency of the vaccine candidate. Then its secondary and tertiary structures, mRNA structure, molecular docking, and immune simulation were predicted using bioinformatics tools. Next, the designed recombinant vaccine construct was synthesized, and cloned into the pET28a vector and expressed in E. coli BL21. Besides, the circular dichroism spectroscopy was utilized for the investigation of the secondary structure changes of the recombinant vaccine construct. The results of the verification assessment of the recombinant vaccine construct expression indicated that in silico analysis was relatively accurate, and relatively change occurred on the protein secondary structure. In our future plan, the vaccine candidate that was confirmed by in silico tools should be validated by further in vitro and in vivo experimental studies.
胶质母细胞瘤是成人原发性恶性脑肿瘤中最常见的一种。目前，手术切除肿瘤，然后辅助放疗和化疗是胶质母细胞瘤的标准治疗方法，但到目前为止还不是有效的治疗方法。因此，开发一种疫苗作为预防或治疗胶质母细胞瘤的安全有效的策略是非常必要的。本研究旨在设计胶质母细胞瘤的多域疫苗。采用计算机方法选择最有效的蛋白结构域，诱导宿主对胶质母细胞瘤产生 B 细胞和 T 细胞免疫反应。IL-13Rα-2（氨基酸位置 27-144）、TNC（氨基酸位置 1900-2100）和 PTPRZ-1（氨基酸位置 731-884）被发现具有很强的诱导免疫反应能力。因此，我们考虑将它们与一个连接子 A(EAAAK)A 融合，构建多结构域重组疫苗。对设计的重组疫苗构建体进行免疫信息学分析，以评估其功效。虽然设计的重组疫苗构建体没有表现出过敏原特性，但它的抗原性估计为 0.78。对重组疫苗构建体的物理化学性质进行了表征，揭示了疫苗候选物的效力。然后，使用生物信息学工具预测其二级和三级结构、mRNA 结构、分子对接和免疫模拟。接下来，合成设计的重组疫苗构建体，并克隆到 pET28a 载体中，在 E. coli BL21 中表达。此外，利用圆二色光谱研究了重组疫苗构建体的二级结构变化。重组疫苗构建体表达验证评估的结果表明，计算机分析相对准确，蛋白质二级结构发生了相对变化。在我们的未来计划中，通过计算机工具确认的候选疫苗应通过进一步的体外和体内实验研究进行验证。
