基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用

刘泽众; 周洁; 朱赟; 陆路; 姜世勃

doi:10.12211/2096-8280.2023-058

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基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用

研究论文 | 更新时间：2025-03-20

- 基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用
- Applications of the recombinant human collagen type Ⅲ-based trimerization motif in the design of vaccines to fight against SARS-CoV-2 and influenza virus
- 合成生物学 2024年5卷第2期页码：385-395
- 作者机构：
  
  1.复旦大学基础医学院，上海 200032
  2.复旦大学药学院药理系，上海 201203
  3.中国科学院生物物理研究所，北京 100101
- 作者简介：
  
  [ "刘泽众（1990—），男，青年研究员，博士生导师。研究方向为微生物学。E-mail：zezhongliu@fudan.edu.cn" ]
  [ "陆路（1982—），男，研究员，博士生导师。研究方向为微生物学。E-mail：lul@fudan.edu.cn" ]
  [ "姜世勃（1953—），男，教授，博士生导师。研究方向为微生物学。E-mail：shibojiang@fudan.edu.cn" ]
- 基金信息：
  
  国家重点研发计划(2022YFC2305800);国家自然科学基金(92169112;82202490)
- DOI：10.12211/2096-8280.2023-058
  中图分类号： R373
- 收稿：2023-08-19，
  
  修回：2023-11-07，
  
  纸质出版：2024-04-30
- 稿件说明：
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刘泽众，周洁，朱赟，陆路，姜世勃. 基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用［J］. 合成生物学， 2024， 5（2）： 385-395

LIU Zezhong， ZHOU Jie， ZHU Yun， LU Lu， JIANG Shibo. Applications of the recombinant human collagen type Ⅲ-based trimerization motif in the design of vaccines to fight against SARS-CoV-2 and influenza virus［J］. Synthetic Biology Journal， 2024， 5（2）： 385-395
刘泽众，周洁，朱赟，陆路，姜世勃. 基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用［J］. 合成生物学， 2024， 5（2）： 385-395 DOI： 10.12211/2096-8280.2023-058.

LIU Zezhong， ZHOU Jie， ZHU Yun， LU Lu， JIANG Shibo. Applications of the recombinant human collagen type Ⅲ-based trimerization motif in the design of vaccines to fight against SARS-CoV-2 and influenza virus［J］. Synthetic Biology Journal， 2024， 5（2）： 385-395 DOI： 10.12211/2096-8280.2023-058.

摘要

新冠病毒等多种包膜病毒的糖蛋白在天然状态下呈现三聚体形式。三聚体蛋白通常较单体蛋白具有更优的免疫原性。目前使用非人蛋白来源的Foldon等三聚体基序可促使目的蛋白形成稳定三聚体，但这些基序的强免疫原性会削弱目的蛋白的免疫应答，限制了其在疫苗抗原设计中的应用。在本研究中，发现并利用重组人源化Ⅲ型胶原蛋白（Rh3C）作为新型三聚体基序，利用合成生物学方法，与新冠病毒刺头（spike）蛋白受体结合域（RBD）融合表达后促使RBD形成三聚体。这种胶原-RBD（Rh3C-RBD）三聚体蛋白在免疫小鼠后，较RBD单体蛋白诱导产生了更高滴度的结合抗体和中和抗体。此外，我们还进一步验证了该Rh3C基序与流感病毒HA1蛋白融合后，较HA1单体蛋白可在小鼠体内诱导更强效的抗体应答。因此，这种新型的Rh3C基序，有望广泛用于三聚体疫苗抗原的设计和优化。

Abstract

Glycoproteins with enveloped viruses

such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)

influenza virus

and human immunodeficiency virus (HIV)

display a trimeric conformation. Different from the monomeric form

the trimeric proteins exhibit superior immunogenicity. Several trimerization motifs

such as Foldon derived from phage T4 fibritin

have been used to promote the formation of trimeric proteins with natural conformations. Although the Foldon-induced trimeric proteins are stable

their high immunogenicity limits applications in the development of vaccine antigens. In a previous study

we developed a recombinant human collagen type Ⅲ protein and determined its crystal structure

revealing a triple-helix conformation. However

the potential of this recombinant protein as a trimerization motif remained unknown. In this study

we demonstrated that the recombinant humanized type Ⅲ collagen (Rh3C) was able to act as a trimerization motif

facilitating the spontaneous trimer formation of the Rh3C-conjugated receptor-binding domain (RBD) within the spike (S) protein of SARS-CoV-2. This trimeric protein could induce a stronger SARS-CoV-2 RBD-specific IgG

IgG1

and IgG2a immune response

when compared with the monomeric RBD protein in the immunized mice. Notably

the Rh3C-RBD protein

when adjuvanted with the novel STING agonist CF501

also elicited significantly higher neutralizing antibody responses against both the pseudotyped SARS-CoV-2 (D614G) and its variant Omicron (BA.2.2) in the immunized mice. To showcase the broad applications of the Rh3C trimerization motif

we further demonstrated that the Rh3C-conjugated HA1 of the influenza virus could also elicit a stronger antibody response than free HA1. Considering the wide distribution of the Rh3C protein in human bodies

its use as a trimerization motif would not induce an immune response due to immune tolerance

thereby allowing the immune response to concentrate on targeted viral proteins. Therefore

this Rh3C-based trimerization motif holds great potential for the design and optimization of vaccines consisting of trimeric protein antigens.

关键词

Keywords

references

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