Hybrid Proteins with Short Conformational Epitopes of the Receptor-Binding Domain of SARS-CoV-2 Spike Protein Promote Production of Virus-Neutralizing Antibodies When Used for Immunization

被引:1
作者
Karyagina, Anna S. [1 ,2 ,3 ]
Gromov, Alexander, V [1 ]
Grunina, Tatyana M. [1 ,2 ]
Lyaschuk, Alexander M. [1 ]
Poponova, Maria S. [1 ]
Kleymenov, Denis A. [1 ]
Strukova, Natalia, V [1 ]
Generalova, Maria S. [1 ]
Ryazanova, Anna, V [1 ]
Galushkina, Zoya M. [1 ]
Dobrynina, Olga Yu [1 ]
Bolshakova, Tatyana N. [1 ]
Sergeeva, Maria, V [4 ]
Romanovskaya-Romanko, Ekaterina A. [4 ]
Krasilnikov, Igor, V [5 ]
Subbotina, Marina E. [1 ,2 ]
Lunin, Vladimir G. [1 ,2 ]
机构
[1] Minist Hlth Russian Federat, Gamaleya Natl Res Ctr Epidemiol & Microbiol, Moscow 123098, Russia
[2] All Russia Res Inst Agr Biotechnol, Moscow 127550, Russia
[3] Lomonosov Moscow State Univ, Belozersky Inst Phys Chem Biol, Moscow 119992, Russia
[4] Minist Hlth Russian Federat, Inst Influenza, St Petersburg 197376, Russia
[5] FMBA, St Petersburg Inst Vaccines & Sera, St Petersburg 198320, Russia
基金
俄罗斯基础研究基金会;
关键词
SARS-CoV-2; S protein; RBD; RBM; epitope vaccine; epitope; aldolase; EOSINOPHILIC INFILTRATION; INFECTION; VACCINES;
D O I
10.1134/S0006297922040022
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Based on the previously developed approach, hybrid recombinant proteins containing short conformational epitopes (a.a. 144-153, 337-346, 414-425, 496-507) of the receptor-binding domain (RBD) of SARS-CoV-2 Spike protein (S protein) were synthesized in Escherichia coli cells as potential components of epitope vaccines. Selected epitopes are involved in protein-protein interactions in the S protein complexes with neutralizing antibodies and ACE2 (angiotensin-converting enzyme 2). The recombinant proteins were used for immunization of mice (three doses with 2-week intervals), and the immunogenicity of protein antigens and ability of the resulting sera to interact with inactivated SARS-CoV-2 and RBD produced in eukaryotic cells were examined. All recombinant proteins showed high immunogenicity; the highest titer in the RBD binding assay was demonstrated by the serum obtained after immunization with the protein containing epitope 414-425. At the same time, the titers of sera obtained against other proteins in the RBD and inactivated virus binding assays were significantly lower than the titers of sera obtained with the previously produced four proteins containing the loop-like epitopes 452-494 and 470-491, the conformation of which was fixed with a disulfide bond. We also studied activation of cell-mediated immunity by the recombinant proteins that was monitored as changes in the levels of cytokines in the splenocytes of immunized mice. The most pronounced increase in the cytokine synthesis was observed in response to the proteins containing epitopes with disulfide bonds (452-494, 470-491), as well as epitopes 414-425 and 496-507. For some recombinant proteins with short conformational epitopes, adjuvant optimization allowed to obtained mouse sera displaying virus-neutralizing activity in the microneutralization assay with live SARS-CoV-2 (hCoV-19/Russia/StPetersburg-3524/2020 EPI_ISL_415710 GISAID). The results obtained can be used to develop epitope vaccines for prevention of COVID-19 and other viral infections.
引用
收藏
页码:319 / 330
页数:12
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