Research Center for High Altitude Medicine, Qinghai University, Kunlun Road, Xining 810001, Qinghai, China; Qinghai-Utah Joint Research Key Lab for High Altitude Medicine, Kunlun Road, Xining 810001, Qinghai, China.
Research Center for High Altitude Medicine, Qinghai University, Kunlun Road, Xining 810001, Qinghai, China; Basic Medicine Department, Qinghai University, Xining 810001, China.
Acta Trop. 2021 Aug;220:105955. doi: 10.1016/j.actatropica.2021.105955. Epub 2021 May 9.
Alveolar echinococcosis is a high-risk parasitic disease caused by the larval stage of Echinococcus multilocularis. The study aimed to predict and identify the dominant Th1/Th2 and B cell epitopes of the antigen protein 14-3-3 beta:alpha from Echinococcus multilocularis.
A comparison of the four amino acid sequences of 14-3-3 beta:alpha was respectively derived from Echinococcus multilocularis, Rattus norvegicus, Canis lupus familiaris, and Homo sapiens was carried out by CLUSTALW to provide a basis for excluding similar epitopes. The amino acid sequence information was analyzed by SOPMA and the homology model was established by Swiss-Model. IEDB and SYFPEITHI were used to predict T cell epitopes. According to the Bcepred and ABCpred, the B cell epitopes were comprehensively predicted and analyzed. The dominant epitopes were validated by Lymphocyte Proliferation, ELISA, ELISpot, and Flow cytometry.
Eight potential epitopes of 14-3-3 from Echinococcus multilocularis were screened according to the results of prediction and analysis: 14-3-3, 14-3-3, 14-3-3, 14-3-3, 14-3-3, 14-3-3, 14-3-3, and 14-3-3. The 3D structure model of the protein was constructed and the location distribution of potential epitope was ascertained. Respectively, the epitopes of the dominant antigen of B cells were validated as 14-3-3 and 14-3-3; the Th1 dominant antigen epitopes were 14-3-3, 14-3-3; and the Th2 dominant epitopes was 14-3-3.
In this study, two dominant antigen epitopes of B cells, two Th1 dominant antigen epitopes, and one Th2 dominant antigen epitope were validated. Our work provides a basis for the subsequent development of efficient and safe vaccines targeting epitopes of Echinococcus multilocularis.
泡型包虫病是一种由细粒棘球绦虫幼虫引起的高风险寄生虫病。本研究旨在预测和鉴定细粒棘球蚴抗原蛋白 14-3-3β:α的优势 Th1/Th2 和 B 细胞表位。
通过 CLUSTALW 比较细粒棘球蚴、挪威鼠、犬和人 14-3-3β:α 的四个氨基酸序列,为排除相似表位提供依据。采用 SOPMA 分析氨基酸序列信息,用 Swiss-Model 建立同源模型。利用 IEDB 和 SYFPEITHI 预测 T 细胞表位。根据 Bcepred 和 ABCpred 综合预测和分析 B 细胞表位。通过淋巴细胞增殖、ELISA、ELISpot 和流式细胞术验证优势表位。
根据预测和分析结果,筛选出细粒棘球蚴 14-3-3β:α 的 8 个潜在表位:14-3-3、14-3-3、14-3-3、14-3-3、14-3-3、14-3-3、14-3-3 和 14-3-3。构建了蛋白质的 3D 结构模型,并确定了潜在表位的位置分布。分别验证了 B 细胞优势抗原的表位为 14-3-3 和 14-3-3;Th1 优势抗原表位为 14-3-3、14-3-3;Th2 优势表位为 14-3-3。
本研究验证了两个 B 细胞优势抗原表位、两个 Th1 优势抗原表位和一个 Th2 优势抗原表位。我们的工作为针对细粒棘球蚴表位的高效、安全疫苗的后续开发提供了依据。