利用免疫信息学方法设计猪产肠毒素大肠杆菌987P-F18蛋白的多表位疫苗

doi:10.7668/hbnxb.20194896

摘要/Abstract

摘要：

利用免疫信息学方法设计一种预防猪大肠杆菌感染的多表位疫苗,阻断产肠毒素大肠杆菌(ETEC)菌株与仔猪肠道上皮细胞的黏附和定植以预防仔猪腹泻。通过免疫信息学工具对ETEC的黏附素987P和F18的B细胞抗原表位、Th细胞表位、CTL细胞表位进行预测,筛选出优势表位。将优势表位按照3种不同表位排列顺序通过Linker 连接成多表位疫苗,并对其抗原性、致敏性以及三级结构进行预测。结果显示,共筛选出7个优势表位。抗原性试验显示,融合多肽Ⅰ的抗原性为1.069 2;融合多肽Ⅱ的抗原性为1.089 2;融合多肽Ⅲ的抗原性为1.050 7,都具有良好的抗原性。过敏性试验结果显示,融合多肽Ⅰ、Ⅱ、Ⅲ都无过敏性。三级结构结果显示,3种融合多肽结构表位的暴露性都较好,容易和抗体结合,并且在蛋白分子的构象中都符合要求。此外,还根据不同的规则,排列了3种氨基酸序列,并按照乳酸菌密码子嗜好性,反向翻译为核苷酸序列,插入至pET28a质粒中,分别获得了pET28a-9F1、pET28a-9F2和 pET28a-9F3质粒。

关键词: 大肠杆菌, 多表位疫苗, 免疫信息学, 987P, F18

Abstract:

A multi-epitope vaccine against porcine Escherichia coli infection was designed by means of immunoinformatics to block the adhesion and colonization of enterotoxigenic Escherichia coli(ETEC)strain to intestinal epithelial cells of piglets to prevent piglet diarrhea.The B cell epitopes,Th cell epitopes,and CTL cell epitopes of adhesin 987P and F18 of ETEC were predicted by immunoinformatics tools,and the dominant epitopes were screened.The dominant epitopes were linked to the multi-epitope vaccine by Linker according to three different epitopes,and their antigenicity,allergenicity,and tertiary structure were predicted.The results showed that seven dominant epitopes were selected.The antigenicity test showed that the antigenicity of fusion peptide Ⅰ,fusion peptide Ⅱ,and fusion peptide Ⅲ were 1.069 2,1.089 2,and 1.050 7,respectively.The results of the allergy test showed that the fusion peptides Ⅰ,Ⅱ,and Ⅲ had no allergen.Based on the three-dimensional modeling of the fusion peptides,the results showed that the structural epitopes were well exposed,easy to bind to antibodies,and met the requirements of epitope design in protein molecular conformation.In addition,according to different rules,we arranged three kinds of amino acid sequences,which were inversely translated into nucleotide sequences according to the codon preference of lactic acid bacteria and inserted into pET28a plasmids to obtain pET28a-9F1,pET28a-9F2,and pET28a-9F3 plasmids respectively.

Key words: Escherichia coli, Multi-epitope vaccine, Immunoinformatics, 987P, F18

中图分类号:

S852.6

徐佳微, 李铁山, 李昕, 刘意, 赵格, 曲志娜, 宋时萍, 张喜悦, 王君玮. 利用免疫信息学方法设计猪产肠毒素大肠杆菌987P-F18蛋白的多表位疫苗[J]. 华北农学报, 2025, 40(S1): 323-328. doi: 10.7668/hbnxb.20194896.

XU Jiawei, LI Tieshan, LI Xin, LIU Yi, ZHAO Ge, QU Zhina, SONG Shiping, ZHANG Xiyue, WANG Junwei. Design of Multi-epitope Vaccine of Porcine Enterotoxigenic Escherichia coli 987P-F18 Protein by Immunoinformatics[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(S1): 323-328. doi: 10.7668/hbnxb.20194896.

http://www.hbnxb.net/CN/Y2025/V40/IS1/323

图/表 11

参考文献 22

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多肽 Polypeptide	表位组成顺序 The composition order of epitopes
融合多肽Ⅰ Fusion peptidesⅠ	B细胞-Th细胞表位、B细胞表位、B细胞-Th细胞-CTL细胞融合表位、B细胞-CTL细胞融合表位、CTL细胞表位
融合多肽Ⅱ Fusion peptidesⅡ	B细胞表位、B细胞-Th细胞表位、CTL细胞表位、B细胞-CTL细胞融合表位、B细胞-Th细胞-CTL细胞融合表位
融合多肽Ⅲ Fusion peptides Ⅲ	CTL细胞表位、B细胞-Th细胞-CTL细胞融合表位、B细胞-CTL细胞融合、B细胞-Th细胞表位、B细胞表位

多肽 Polypeptide	表位组成顺序 The composition order of epitopes
融合多肽Ⅰ Fusion peptidesⅠ	B细胞-Th细胞表位、B细胞表位、B细胞-Th细胞-CTL细胞融合表位、B细胞-CTL细胞融合表位、CTL细胞表位
融合多肽Ⅱ Fusion peptidesⅡ	B细胞表位、B细胞-Th细胞表位、CTL细胞表位、B细胞-CTL细胞融合表位、B细胞-Th细胞-CTL细胞融合表位
融合多肽Ⅲ Fusion peptides Ⅲ	CTL细胞表位、B细胞-Th细胞-CTL细胞融合表位、B细胞-CTL细胞融合、B细胞-Th细胞表位、B细胞表位

序号 Number	起始位点 Starting site	终止位点 Termination site	肽段 Peptide	长度/aa Length
1	20	29	PEVNGNRTST	10
2	36	88	AISGHGTVVDFKLKPAPGSNDCLAKTNARIDWSGSMNSLGFNNTASGNTAAKG	53
3	96	114	TNVGNGSGGANINTSFTTA	19
4	118	126	HTSAIQSFN	9

序号 Number	起始位点 Starting site	终止位点 Termination site	肽段 Peptide	长度/aa Length
1	20	29	PEVNGNRTST	10
2	36	88	AISGHGTVVDFKLKPAPGSNDCLAKTNARIDWSGSMNSLGFNNTASGNTAAKG	53
3	96	114	TNVGNGSGGANINTSFTTA	19
4	118	126	HTSAIQSFN	9

序号 Number	起始位点 Starting site	肽段 Peptide	评分 Scoring
1	35	AAISGHGTVVDFKLKP	0.93
2	41	GTVVDFKLKPAPGSND	0.88
3	9	GKITSATCTIDPEVNG	0.86
4	62	NARIDWSGSMNSLGFN	0.85
5	15	TCTIDPEVNGNRTSTI	0.84
6	129	AQLKKDDRAPSNGGYK	0.84
7	27	TSTIDLGQAAISGHGT	0.83
8	82	GNTAAKGYHMTLRATN	0.80
9	2	TGTINFNGKITSATCT	0.79

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