Institute of Molecular Biology and Biotechnology, Bahauddin Zakariya University, Multan, Pakistan.
School of Information and Technology, Wenzhou Business College, Wenzhou, Zhejiang, China.
Biomed Res Int. 2022 May 31;2022:4792374. doi: 10.1155/2022/4792374. eCollection 2022.
Liver cancer (LC) is the most devastating disease affecting a large set of populations in the world. The mortality due to LC is escalating, indicating the lack of effective therapeutic options. Immunotherapeutic agents may play an important role against cancer cells. As immune cells, especially T lymphocytes, which are part of cancer immunology, the design of vaccine candidates for cytotoxic T lymphocytes may be an effective strategy for curing liver cancer.
In our study, based on an immunoinformatics approach, we predicted potential T cell epitopes of MHC class I molecules using integrated steps of data retrieval, screening of antigenic proteins, functional analysis, peptide synthesis, and experimental investigations. We predicted the binding affinity of epitopes LLECADDRADLAKY, VSEHRIQDKDGLFY, and EYILSLEELVNGMY of LC membrane-bounded extracellular proteins including butyrophilin-like protein-2 (BTNL2), glypican-3 (GPC3), and serum albumin (ALB), respectively, with MHC class I molecules (allele: HLA-A∗01:01). These T cell epitopes rely on the level of their binding energy and antigenic properties. We designed and constructed a trivalent immunogenic model by conjugating these epitopes with linkers to activate cytotoxic T cells. For validation, the nonspecific hematological assays showed a significant rise in the count of white blood cells (5 × 10/l), lymphocytes (13 × 10/l), and granulocytes (5 × 10/l) compared to the control after administration of trivalent peptides. Specific immunoassays including granzyme B and IgG ELISA exhibited the significant concentration of these effector molecules in blood serum, indicating the activity of cytotoxic T cells. Granzyme concentration increased to 1050 pg/ml at the second booster dose compared to the control (95 pg/ml), while the concentration of IgG raised to 6 g/l compared to the control (2 g/l).
We concluded that a potential therapeutic trivalent vaccine can activate and modulate the immune system to cure liver cancer on the basis of significant outcomes of specific and nonspecific assays.
肝癌（LC）是全球影响大量人群的最具破坏性疾病。由于 LC 导致的死亡率不断上升，表明缺乏有效的治疗选择。免疫治疗药物可能在对抗癌细胞方面发挥重要作用。作为免疫细胞，尤其是 T 淋巴细胞，它们是癌症免疫学的一部分，设计针对细胞毒性 T 淋巴细胞的疫苗候选物可能是治愈肝癌的有效策略。
在我们的研究中，基于免疫信息学方法，我们使用数据检索、抗原蛋白筛选、功能分析、肽合成和实验研究等综合步骤，预测了 MHC Ⅰ类分子的潜在 T 细胞表位。我们预测了 LC 膜结合细胞外蛋白（包括丁酰膦蛋白样蛋白 2（BTNL2）、聚糖蛋白 3（GPC3）和血清白蛋白（ALB））中的表位 LLECADDRADLAKY、VSEHRIQDKDGLFY 和 EYILSLEELVNGMY 与 MHC Ⅰ类分子（等位基因：HLA-A*01:01）的结合亲和力。这些 T 细胞表位依赖于它们的结合能和抗原性。我们通过连接子将这些表位与连接子偶联，设计并构建了一种三价免疫原性模型，以激活细胞毒性 T 细胞。为了验证，非特异性血液学检测显示，与对照组相比，给药后白细胞（5×10/l）、淋巴细胞（13×10/l）和粒细胞（5×10/l）计数显著升高。与对照组（95pg/ml）相比，特异性免疫测定（包括颗粒酶 B 和 IgG ELISA）显示血液中这些效应分子的浓度显著升高，表明细胞毒性 T 细胞的活性。与对照组（95pg/ml）相比，第二次增强剂量时颗粒酶浓度增加到 1050pg/ml，而 IgG 浓度增加到 6g/l 与对照组（2g/l）相比。
我们的结论是，一种潜在的治疗性三价疫苗可以在特定和非特异性检测的显著结果的基础上激活和调节免疫系统，从而治愈肝癌。
