Antibodies against VP3 protein of echovirus 30 (Picornaviridae: Enterovirus: Enterovirus betacoxsackie) neutralize virus in vitro

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Abstract

Introduction. The widespread occurrence of enterovirus echovirus 30 (E30) and cases of severe disease indicate the need for vaccine development. Epitopes for neutralizing antibodies and T-cell response have been found in VP3 proteins of some enteroviruses. However, the immunogenic properties of VP3 E30 have not been studied. The aim of this work is to characterize the immunogenic properties of VP3 E30 and testing the virus-neutralizing properties of antibodies against VP3 E30.

Materials and methods. VP3E30 and the chimeric protein SN-VP3E30, consisting of the S region of norovirus VP1 and VP3 E30 were expressed in Escherichia coli. SN-VP3E30 was used to form virus-like particles (VLPs). The effect on human dendritic cells (DCs) was assessed by flow cytometry to measure changes in HLA-DR, CCR7, CD80, CD83 and CD86 expression. BALB/c mice and guinea pig were used for immunization with SN-VP3E30. Antibody titers and avidity were determined by ELISA. The interaction of antibodies with VP3 E30 was studied by immunoelectron microscopy and virus neutralization in embryonal rhabdomyosarcoma (RD) cell culture.

Results. Recombinant VP3 E30 caused incomplete DCs maturation, characterized by the understimulation of the CCR7 chemokine receptor expression. Inclusion of VP3 in the chimeric VLPs resulted in complete DCs maturation and a strong humoral immune response in laboratory animals. Antibodies against VP3 were characterized by high avidity, the ability to induce agglomeration of viral particles and neutralization of E30 in RD cell culture.

Conclusion. The obtained results indicate that VP3 can be used as an antigen in the composition of a subunit vaccine against enterovirus E30.

About the authors

Dmitry V. Novikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); N.I. Lobachevsky State University of Nizhny Novgorod

Author for correspondence.
Email: novikov.dv75@mail.ru
ORCID iD: 0000-0001-7049-6935

PhD, Leading Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950; Nizhny Novgorod, 603950

Dmitry A. Melentev

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); N.I. Lobachevsky State University of Nizhny Novgorod

Email: dim-melente@yandex.ru
ORCID iD: 0000-0002-2441-6874

Junior Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950; Nizhny Novgorod, 603950

Vladimir Yu. Talayev

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: talaev@inbox.ru
ORCID iD: 0000-0003-1993-0622

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

Russian Federation, Nizhny Novgorod, 603950

Nadezhda A. Novikova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: novikova_na@mail.ru
ORCID iD: 0000-0002-3710-6648

Professor. Head of the laboratory of molecular epidemiology of viral infections

Russian Federation, Nizhny Novgorod, 603950

Ekaterina V. Mokhonova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: ekaterinamohonova@yandex.ru
ORCID iD: 0000-0002-9742-7646

Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950

Alexander U. Kashnikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: mevirfc@mail.ru
ORCID iD: 0000-0003-1033-7347

Researcher, laboratory of molecular epidemiology of viral infections

Russian Federation, Nizhny Novgorod, 603950

Irina E. Zaichenko

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: imm.irina@mail.ru
ORCID iD: 0000-0001-5063-3111

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

Russian Federation, Nizhny Novgorod, 603950

Marya V. Svetlova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: marya.talaeva@yandex.ru
ORCID iD: 0000-0003-4097-6780

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

Russian Federation, Nizhny Novgorod, 603950

Elena V. Kurkova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: el2v@mail.ru
ORCID iD: 0000-0003-1801-9693

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

Russian Federation, Nizhny Novgorod, 603950

Olga N. Babaykina

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: olga_babaykina@inbox.ru
ORCID iD: 0000-0003-4527-6134

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

Russian Federation, Nizhny Novgorod, 603950

Vladislav A. Lapin

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); N.I. Lobachevsky State University of Nizhny Novgorod

Email: fridens.95@yandex.ru
ORCID iD: 0000-0002-5905-5722

Junior Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950; Nizhny Novgorod, 603950

Maria I. Tsyganova

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)

Email: maria_che@mail.ru
ORCID iD: 0000-0002-2811-6844

PhD, Leading Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950

Dmitry E. Zaitsev

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); N.I. Lobachevsky State University of Nizhny Novgorod

Email: mitya.zaitseff@yandex.ru
ORCID iD: 0000-0002-7663-6924

Junior Researcher, laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950; Nizhny Novgorod, 603950

Viktor V. Novikov

Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); N.I. Lobachevsky State University of Nizhny Novgorod

Email: mbre@mail.ru
ORCID iD: 0000-0002-2449-7213

DSc (Biology), Professor, Leading Researcher, Head of the laboratory of immunochemistry

Russian Federation, Nizhny Novgorod, 603950; Nizhny Novgorod, 603950

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2. Fig. 1. The SN-VP3E30 protein forms virus-like particles (VLPs). a – electrophoretic mobility of purified SN-VP3E30 (1) compared to PageRuler Plus Prestained Protein Ladder (Thermo Scientific) (2); b – electron micrograph of VLP formed by e SN-VP3E30 (12,000× magnification).

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3. Fig. 2. Expression of HLA-DR, CD80, CD83, CD86, and CCR7 molecules on dendritic cells (DCs) after incubation with VP3 or SN-VP3E30. a – the fluorescence intensity (GMFI); b – the percentage of cells bearing the marker (Tukey histograms). The cell type, their stimulators, and analyzed markers are shown below the histograms. Differences from unstimulated DCs are indicated: † – p < 0.05, †† – p < 0.005; differences from DCs incubated with negative control are indicated: * – p < 0.05, ** – p < 0.005 (paired Wilcoxon signed rank test).

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4. Fig. 3. Characteristics of antibodies to VP3E30 after mice immunization by the chimeric protein SN-VP3E30. a – antibody titers; b – antibody avidity index. 1 – immunization without adjuvant (n = 10), 2 – immunization with adjuvant (n = 10).

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5. Fig. 4. Immunoelectron micrographs of E30 (12,000× magnification). a – preparation with antibodies against norovirus VP1; b – Е30-еС2 1194/24 aggregates after interaction with antibodies against VP3E30; с – Е30-h 2671/17 aggregates after interaction with antibodies against VP3E30.

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Copyright (c) 2025 Novikov D.V., Melentev D.A., Talayev V.Y., Novikova N.A., Mokhonova E.V., Kashnikov A.U., Zaichenko I.E., Svetlova M.V., Kurkova E.V., Babaykina O.N., Lapin V.A., Tsyganova M.I., Zaitsev D.E., Novikov V.V.

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