Expression of dengue capsid-like particles in silkworm and display of envelope domain III of dengue virus serotype 2

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2024-07-04 DOI:10.1016/j.pep.2024.106543

Krishna Raja Muthuraman , Doddy Irawan Setyo Utomo , Mami Matsuda , Ryosuke Suzuki , Enoch Y. Park

{"title":"Expression of dengue capsid-like particles in silkworm and display of envelope domain III of dengue virus serotype 2","authors":"Krishna Raja Muthuraman , Doddy Irawan Setyo Utomo , Mami Matsuda , Ryosuke Suzuki , Enoch Y. Park","doi":"10.1016/j.pep.2024.106543","DOIUrl":null,"url":null,"abstract":"<div><p>Dengue virus (DENV) is a considerable public health threat affecting millions of people globally. Vaccines for dengue are an important strategy to reduce the disease burden. We expressed capsid (C2) and envelope domain III of dengue virus serotype 2 (2EDIII) separately in the silkworm expression system. We conjugated them employing the monomeric streptavidin (mSA2) and biotin affinity to display the antigenic 2EDIII on the C2-forming capsid-like particle (CLP). Purified 2EDIII-displaying C2 (CLP/2EDIII) was immunogenic in BALB/c mice, eliciting neutralizing antibodies confirmed by a single-round infectious particle (SRIP) neutralization assay. Th1 cytokine levels were upregulated for the CLP/2EDIII group, and the anti-inflammatory IL-10 and pro-inflammatory IL-6 cytokine levels were also raised compared to the 2EDIII and the control groups. Elevated cytokine levels for CLP/2EDIII indicate the importance of displaying the 2EDIII as CLP/2EDIII rather than as an individual subunit. This study is the first to express the C2 protein as self-assembling CLP <em>in vivo</em> and 2EDIII separately in the silkworm expression system and conjugate them to form a monovalent CLP. Thus, this CLP/2EDIII display method may pave the way for an efficient tetravalent dengue vaccine candidate.</p></div>","PeriodicalId":20757,"journal":{"name":"Protein expression and purification","volume":"222 ","pages":"Article 106543"},"PeriodicalIF":1.4000,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Protein expression and purification","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1046592824001153","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

Abstract

Dengue virus (DENV) is a considerable public health threat affecting millions of people globally. Vaccines for dengue are an important strategy to reduce the disease burden. We expressed capsid (C2) and envelope domain III of dengue virus serotype 2 (2EDIII) separately in the silkworm expression system. We conjugated them employing the monomeric streptavidin (mSA2) and biotin affinity to display the antigenic 2EDIII on the C2-forming capsid-like particle (CLP). Purified 2EDIII-displaying C2 (CLP/2EDIII) was immunogenic in BALB/c mice, eliciting neutralizing antibodies confirmed by a single-round infectious particle (SRIP) neutralization assay. Th1 cytokine levels were upregulated for the CLP/2EDIII group, and the anti-inflammatory IL-10 and pro-inflammatory IL-6 cytokine levels were also raised compared to the 2EDIII and the control groups. Elevated cytokine levels for CLP/2EDIII indicate the importance of displaying the 2EDIII as CLP/2EDIII rather than as an individual subunit. This study is the first to express the C2 protein as self-assembling CLP in vivo and 2EDIII separately in the silkworm expression system and conjugate them to form a monovalent CLP. Thus, this CLP/2EDIII display method may pave the way for an efficient tetravalent dengue vaccine candidate.

查看原文本刊更多论文

在家蚕体内表达登革热包膜样颗粒并显示登革热病毒血清 2 型的包膜结构域 III。

登革热病毒（DENV）是影响全球数百万人的严重公共卫生威胁。登革热疫苗是减轻疾病负担的重要策略。我们在家蚕表达系统中分别表达了登革病毒血清型 2（2EDIII）的囊膜（C2）和包膜结构域 III。我们利用单体链霉亲和素（mSA2）和生物素的亲和力将它们共轭，在C2形成的类囊体颗粒（CLP）上显示抗原性2EDIII。纯化的显示2EDIII的C2（CLP/2EDIII）在BALB/c小鼠中具有免疫原性，通过单轮感染性颗粒（SRIP）中和试验证实可激发中和抗体。与 2EDIII 组和对照组相比，CLP/2EDIII 组的 Th1 细胞因子水平升高，抗炎性 IL-10 和促炎性 IL-6 细胞因子水平也升高。CLP/2EDIII细胞因子水平的升高表明，将2EDIII作为CLP/2EDIII而不是单独的亚单位显示非常重要。本研究首次在家蚕表达系统中将 C2 蛋白以自组装 CLP 的形式在体内表达，并将 2EDIII 单独表达，然后将它们共轭形成单价 CLP。因此，这种CLP/2EDIII展示方法可能会为高效的四价登革热候选疫苗铺平道路。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.