Intranasal vaccine against COVID-19 based on a recombinant variant of the Sendai virus (Paramyxoviridae: Respirovirus) strain Moscow

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Abstract

Introduction. Intranasal vaccination using live vector vaccines based on non-pathogenic or slightly pathogenic viruses is the one of the most convenient, safe and effective ways to prevent respiratory infections, including COVID-19. Sendai virus is the best suited for this purpose, since it is respiratory virus and is capable of limited replication in human bronchial epithelial cells without causing disease.

The aim of the work is to design and study the vaccine properties of recombinant Sendai virus, Moscow strain, expressing secreted receptor-binding domain of SARS-CoV-2 Delta strain S protein (RBDdelta) during a single intranasal immunization.

Materials and methods. Recombinant Sendai virus carrying insertion of RBDdelta transgene between P and M genes was constructed using reverse genetics and synthetic biology methods. Expression of RBDdelta was analyzed by Western blot. Vaccine properties were studied in two models: Syrian hamsters and BALB/c mice. Immunogenicity was evaluated by ELISA and virus-neutralization assays. Protectiveness was assessed by quantitation of SARS-CoV-2 RNA in RT-PCR and histological analysis of the lungs.

Results. Based on Sendai virus Moscow strain, a recombinant Sen-RBDdelta(M) was constructed that expressed a secreted RBDdelta immunologically identical to natural SARS-CoV-2 protein. A single intranasal administration of Sen-RBDdelta(M) to hamsters and mice significantly, by 15 and 107 times, respectively, reduced replicative activity of SARS-CoV-2 in lungs of animals, preventing the development of pneumonia. An effective induction of virus-neutralizing antibodies has also been demonstrated in mice.

Conclusion. Sen-RBDdelta(M) is a promising vaccine construct against SARS-CoV-2 infection and has a protective properties even after a single intranasal introduction.

About the authors

Gleb A. Kudrov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: kudrov_ga@vector.nsc.ru
ORCID iD: 0000-0002-8251-7040

junior research assistant

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Sergei S. Zainutdinov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: zaynutdinov_ss@vector.nsc.ru
ORCID iD: 0000-0001-5818-4402

researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Antonina A. Grazhdantseva

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: gaa@vector.nsc.ru
ORCID iD: 0000-0001-7712-3699

PhD, senior researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Andrey V. Shipovalov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: shipovalov_av@vector.nsc.ru
ORCID iD: 0000-0003-1201-8307

researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Galina F. Sivolobova

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: sgf@vector.nsc.ru
ORCID iD: 0000-0002-8362-0314

PhD, senior researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Anastasiya V. Semenova

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: tkacheva_av@vector.nsc.ru
ORCID iD: 0000-0001-7767-0537

PhD, senior researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Iuliia A. Merkuleva

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: merkuleva_yua@vector.nsc.ru
ORCID iD: 0000-0002-6974-0686

PhD, junior research assistant

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Dmitry N. Shcherbakov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: scherbakov_dn@vector.nsc.ru
ORCID iD: 0000-0001-8023-4453

PhD, head of laboratory

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Oleg S. Taranov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: taranov@vector.nsc.ru
ORCID iD: 0000-0002-6746-8092

head of department

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Anna V. Zaykovskaya

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: zaykovskaya_av@vector.nsc.ru
ORCID iD: 0000-0002-0450-5212

PhD, senior researcher

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Irina S. Shulgina

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: Shulgina_is@vector.nsc.ru
ORCID iD: 0000-0002-6850-338X

graduate student

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Oleg V. Pyankov

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Email: pyankov@vector.nsc.ru
ORCID iD: 0000-0003-3340-8750

PhD, head of department

Russian Federation, 630559, Koltsovo, Novosibirsk Region

Galina V. Kochneva

State Research Center of Virology and Biotechnology “Vector” of Rospotrebnadzor

Author for correspondence.
Email: kochneva@vector.nsc.ru
ORCID iD: 0000-0002-2420-0483

Dr. Sci. (Biol.), head of laboratory

Russian Federation, 630559, Koltsovo, Novosibirsk Region

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2. Fig. 1. Scheme for constructing the recombinant Sen-RBDdelta(M) variant of the Sendai virus strain Moscow with the introduction of the RBDdelta transgene between the P and M genes. Synthesis of genomic RNA of recombinant Sendai virus strains is carried out from plasmid DNA under the control of T7 phage polymerase (indicated by the arrow). рT7 – promoter; TT7 – T7 polymerase terminator; rib – ribozyme.

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3. Fig. 2. Western blot analysis of RBDdelta expression by recombinant Sen-RBDdelta(M) virus: a, c – in allantoic fluid of 11-day-old chicken embryos; b – in culture medium of LLC-MK2 cells. Polyclonal blood serum of a mouse immunized by RBDdelta, dilution 1 : 500 (a, b) or blood serum of vaccinated convalescent COVID-19, dilution 1 : 200 (c) was used as primary antibodies. M – molecular weight control; 1 – negative control of the allantoic fluid of uninfected chicken embryos (a, c) or the culture medium of uninfected LLC-MK2 cells (b); 2 – positive control of RBD (200 ng, 35 kDa) expressed in CHO-K1 cells; 3 – empty track; 4 – allantoic fluid of chicken embryos infected with Sen-RBDdelta(M), 15 µl (a, c), or culture medium of cells infected with Sen-RBDdelta(M) (b).

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4. Fig. 3. Clinical disease in hamsters after intranasal infection with SARS-CoV-2 delta strain: a – dynamics of weight change, points are median values, vertical lines are 95% confidence intervals; b – viral load (copies/ml) in nasal cavity flushes; c – viral load (copies/mg) in homogenates of nasal cavity and lung tissues, dots are individual values, horizontal lines are medians, vertical lines are 95% confidence intervals. p-values are indicated for statistically significant differences (p < 0.05) in the Mann–Whitney U-test.

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5. Fig. 4. Histological section of a Syrian hamster lung from the control (a) and the vaccinated group (b).

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6. Fig. 5. Total and neutralizing IgG antibodies against the SARS-CoV-2 delta strain in the sera of immunized BALB/c mice: a – optical density value (OD450) determined by ELISA; dots – individual values, histogram tops – medians, vertical lines – 95% confidence intervals; b – neutralization titers (NT50), points – individual values, histogram tops – geometric means, vertical lines – standard deviations; horizontal dotted line – threshold value (NT50 < 1 : 10). p-values are indicated for statistically significant differences (p < 0.05) in Dunn’s Multiple Comparison Test.

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7. Fig. 6. Viral load (copies/mg) in nasal cavity and lung tissue homogenates of BALB/c mice on day 5 after infection with Gamma strain SARS-CoV-2. In the diagram, the points are individual values, the horizontal lines are medians, and the vertical lines are 95% confidence intervals. P-values are indicated for statistically significant differences (p < 0.05) in Dunn’s Multiple Comparison Test.

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Copyright (c) 2023 Kudrov G.A., Zainutdinov S.S., Grazhdantseva A.A., Shipovalov A.V., Sivolobova G.F., Semenova A.V., Merkuleva I.A., Shcherbakov D.N., Taranov O.S., Zaykovskaya A.V., Shulgina I.S., Pyankov O.V., Kochneva G.V.

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