Coronavirus spike protein fragment-containing chimeric virus-like particles stimulate human dendritic cell maturation

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Abstract

Introduction. Viral capsid proteins can assemble into virus-like particles lacking infectivity and bearing parental virus antigens or artificially introduced antigens from other pathogens. At least some of such particles are highly immunogenic and could serve as a platform for promising vaccines. In this work, we assessed an effect of virus-like particles decorated with a SARS-CoV-2 spike protein fragment on human dendritic cell phenotype and functional properties. Materials and methods. The virus-like particles were assembled using chimeric molecules obtained by fusing genetic sequences encoding a norovirus major capsid protein VP1 fragment and a coronavirus spike protein fragment, including the receptor-binding domain. Dendritic cells were obtained from monocytes in vitro. Results. Incubation of immature dendritic cells with virus-like particles induced their phenotypic and functional maturation. The former was revealed by significantly increased expression of HLA-DR, CD80, CD86 and CD83. Dendritic cell phenotype after incubation with virus-like particles at the maximum concentration of 10 μg/ml did not differ significantly from that of mature dendritic cells in positive control. Along with phenotypic maturation, virus-like particles caused a manifold increase in the production of pro-inflammatory tumor necrosis factor-α, anti-inflammatory interleukin-10, as well as interleukin-6, which can stimulate both antibody synthesis and cellular pro-inflammatory reactions. The pronounced stimulation of dendritic cells by virus-like particles coated with coronavirus antigens evidence about successful particle recognition. Finally, we discuss plausible mechanisms for recognition of such virus-like particles by dendritic cell receptors. Conclusion. It has been shown that chimeric virus-like particles induced phenotypic and functional dendritic cell maturation, which is manifested by markedly elevated expression of functionally important membrane molecules, as well as a manifold rise in production of cytokines with a wide functional range. In our opinion, the data obtained indicate a promise of using virus-like particles based on norovirus proteins to display SARS-CoV-2 antigens.

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Chimeric virus-like particles containing a fragment of the coronavirus spike protein stimulate the maturation of human dendritic cells

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About the authors

V. Yu. Talayev

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Author for correspondence.
Email: talaev@inbox.ru

DSc (Medicine), Professor, Head of the Laboratory of Cellular Immunology

Россия, Nizhniy Novgorod

D. V. Novikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

PhD (Biology), Leading Researcher, Laboratory of Immunochemistry

Россия, Nizhniy Novgorod

I. Ye. Zaichenko

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

PhD (Biology), Leading Researcher, Laboratory of Cellular Immunology

Россия, Nizhniy Novgorod

M. V. Svetlova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

PhD (Biology), Senior Researcher, Laboratory of Cellular Immunology

Россия, Nizhniy Novgorod

E. V. Voronina

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

PhD (Biology), Senior Researcher, Laboratory of Cellular Immunology

Россия, Nizhniy Novgorod

O. N. Babaykina

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

PhD (Medicine), Senior Researcher, Laboratory of Cellular Immunology

Россия, Nizhniy Novgorod

V. A. Lapin

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

Junior Researcher, Laboratory of Immunochemistry

Россия, Nizhniy Novgorod

D. A. Melentiev

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

Junior Researcher, Laboratory of Immunochemistry

Россия, Nizhniy Novgorod

N. A. Novikova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

DSc (Biology), Professor, Laboratory of Molecular Epidemiology of Viral Infections

 

Россия, Nizhniy Novgorod

A. Yu. Kashnikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

Researcher, Laboratory of Molecular Epidemiology of Viral Infections

Россия, Nizhniy Novgorod

V. V. Novikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing

Email: talaev@inbox.ru

DSc (Biology), Professor, Head of the Laboratory of Immunochemistry

Россия, Nizhniy Novgorod

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2. Figure 1. Characteristics of SN-RBD VLPs. Note. A) A scheme of the genetic construct encoding the chimeric protein SN-RBD. B) Electropherogram of purified SN-RBD protein. С) VLPs electron microscopic images; magnification ×15 000.

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3. Figure 2. Expression of membrane molecules on unstimulated iDCs (thin gray line) and DCs incubated with 10 μg/ml SN-RBD VLPs (thick black line) or equivalent CS amount (thin black line). Note. The dotted line shows DC negative control staining exposed to VLPs. Membrane molecules and fluorescence intensity are indicated below the x-axis. A representative experiment (n = 11) is presented.

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4. Figure 3. Effect of SN-RBD VLPs on DC maturation. Note. The y-axis: percentage of cells expressing the molecule or the GMFI for stained molecule. The type of stimulant is indicated under the x-axis, the VLP concentration is indicated in the legend. Significant differences (p < 0.05 in paired t-test with Bonferroni correction) compared to unstimulated DCs (*) and CS-treated DCs (†). Data are presented as M±m (n = 11).

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5. Figure 4. Effect of SN-RBD VLPs on cytokine production in DC cultures. Note. The type of stimulant is under the x-axis, cytokine concentrations are shown on the y-axis, VLP concentration shown in the legend. Significant differences (p < 0.05; Wilcoxon test with Bonferroni correction) when compared with unstimulated DCs (*) and DCs cultured with CS (†). Data are presented as Median ± quartile (n = 11).

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Copyright (c) 2024 Talayev V.Y., Novikov D.V., Zaichenko I.Y., Svetlova M.V., Voronina E.V., Babaykina O.N., Lapin V.A., Melentiev D.A., Novikova N.A., Kashnikov A.Y., Novikov V.V.

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