| 金亚珉,杨姣姣,张 婷,王志荣,许雪梅.B/Yamagata系流感病毒血凝素（HA）蛋白疫苗的筛选研究[J].,2024,(15):2801-2804 |
| B/Yamagata系流感病毒血凝素（HA）蛋白疫苗的筛选研究 |
| Screening of Hemagglutinin (HA) Protein Vaccine for Influenza Viruses B/Yamagata Lineage |
| 投稿时间：2024-04-01 修订日期：2024-04-17 |
| DOI：10.13241/j.cnki.pmb.2024.15.001 |
| 中文关键词: B/Yamagata 血凝素（HA） 重组蛋白疫苗 |
| 英文关键词: B/Yamagata Hemagglutinin (HA) Recombinant protein vaccines |
| 基金项目:中国医学科学院医学与健康科技创新工程项目基金（2021-I2M-1-043） |
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| 中文摘要: |
| 摘要 目的：筛选表达量高且具有血凝抑制活性的B/Yamagata系流感病毒血凝素（HA）重组蛋白。方法：以B/Singapore/INFTT-16-0610/2016的HA为基础，在HA胞外区C末端分别融合GCN4pLL三聚化基序、GCN4pLL-半胱氨酸及GCN4pLL-七鳃鳗可变淋巴细胞受体-B（VLR-B）抗体C端的肽段序列，分别获得HA突变体基因BY-T、BY-LLc及BY-PLc；分别插入pFastBac1，采用杆状病毒昆虫细胞表达系统进行表达。表达鉴定后，Strep-Tactin亲和层析纯化，然后鉴定纯化突变体蛋白的寡聚程度及血凝活性。突变体蛋白免疫小鼠，检测免疫血清HA特异性IgG抗体和血凝抑制（HAI）抗体水平。结果：各HA突变体均能够有效表达，BY-T突变体表达水平最高，易于纯化，纯度好，呈多聚化形式存在；BY-LLc及BY-PLc表达水平次之，呈高聚化形式存在。三种HA突变体蛋白的血凝活性相当，均可有效诱发的HA特异性结合的IgG抗体和HAI抗体。结论：HA蛋白胞外区C端融合寡聚化基序GCN4pLL的突变体易于纯化，具有血凝活性，可诱发小鼠产生HA特异性抗体和HAI抗体。研究为B/Yamagata系流感病毒重组蛋白疫苗的研发策略提供了参考。 |
| 英文摘要: |
| ABSTRACT Objective: To screen the recombinant hemagglutinin (HA) protein with high expression level and hemagglutinin inhibitory activity for B/Yamagata influenza virus. Methods: Based on the HA of B/Singapore/INFTT-16-0610/2016, the peptide sequences of GCN4pLL trimerization motif, GCN4pLL-cysteine, and GCN4pLL-the sequences from C-terminus of lamprey variable lymphocyte receptor (VLR)-B antibodies were fused to C-terminus of HA ectodomain, to obtain HA mutant genes BY-T, BY-LLc, and BY-PLc. They were inserted into pFastBac1 separately and expressed using baculovirus expression vector system. After expression identification, Strep-Tactin affinity chromatography purification was performed to purify the mutant proteins followed by identification of the degree of oligomerization and hemagglutination activity of the purified mutant proteins. Anti-HA IgG titer and hemagglutination inhibition (HAI) antibody titer in sera of mice immunized with HA mutants were detected. Results: All HA mutants were effectively expressed. The BY-T mutant had the highest expression level, was easy to purify, had good purity, and existed in a polymeric form. The expression levels of BY-LLc and BY-PLc are secondary and exist in a highly polymerization form. The hemagglutination activities of the three HA mutant proteins were comparable, and all of them could effectively induce anti-HA IgG and HAI antibodies. Conclusion: The HA protein ectodomain mutant fused with oligomeric motif GCN4pLL at the C-terminus is easy to purify and has hemagglutination activity, which can induce anti-HA IgG and HAI antibodies in mice. The results provide a reference for the development strategy of recombinant protein vaccines against influenza virus B/Yamagata lineage. |
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