Immunization of mice with pVAXrbd DNA vaccine by jet injection induces a stronger immune response and protection against SARS-CoV-2 compared to intramuscular injection via syringe

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During the COVID-19 pandemic, it became clear that to ensure global health security, it is essential to have a developed platform that can be used to quickly develop a safe, low-cost, effective vaccine. DNA vaccines have several advantages over other platforms, including rapid development and ease of production. They are more stable than mRNA vaccines. Unlike viral vector-based vaccines, DNA vaccines do not induce anti-vector immunity. One of the disadvantages of DNA vaccines is their relatively low immunogenicity. This problem can be solved using jet injection. Here, we evaluated and confirmed the efficiency of an inexpensive, simple, and safe method for delivering the naked DNA vaccine pVAXrbd encoding the receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein using a spring-loaded jet injector. Based on the results of histological analysis, optimal condition, were determined that ensure low tissue trauma in laboratory animals upon administration of pVAXrbd. An optimized immunization protocol for BALB/c mice was used to compare the immunogenicity of pVAXrbd with two different administration schemes: using a jet injector under the skin and into the adjacent muscle layers or intramuscularly using a syringe with a needle. Mice immunized with ‘naked’ pVAX-rbd were shown to produce high levels of specific virus-neutralizing antibodies. The vaccine also induced a strong RBD-specific T-cell response. As shown by quantitative PCR analysis of viral RNA, vaccinated mice infected with the Gamma variant of SARS-CoV-2 developed a protective immune response; moreover, it was more pronounced in animals to which the DNA-vaccine was administered using a jet injector compared to those immunized intramuscularly. Thus, the introduction of a DNA-vaccine using jet injection effectively activates both types of the immune response and leads to a decrease in the viral load. Jet injection is a promising method for delivering DNA vaccines, characterized by low cost, simplicity, technological administration and minimal pain for the patient.

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作者简介

D. Kisakov

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

编辑信件的主要联系方式.
Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

M. Borgoyakova

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

L. Kisakova

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

E. Starostina

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

O. Pyankov

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

А. Zaykovskaya

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

O. Taranov

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

E. Ivleva

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

N. Rudometova

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

V. Yakovlev

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

Е. Tigeeva

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

M. Azaev

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

I. Belyakov

Moscow Financial-Industrial University “Synergy”

Email: def_2003@mail.ru
俄罗斯联邦, Moscow, 129090

A. Rudometov

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

A. Ilyichev

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

L. Karpenko

State Scientific Center of Virology and Biotechnology “Vector”, Rospotrebnadzor

Email: def_2003@mail.ru

Center for Genomic Research of the World Level of Biosafety and Technological Independence, Federal Scientific and Technical Program for Development of Genetic Technologies

俄罗斯联邦, Koltsovo, Novosibirsk Region, 630559

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2. Fig. 1. Hematoxylin and eosin stained tissue samples of the quadriceps femoris muscle of the left hind paw of mice from different groups. Tissue samples of 1 mm in size were taken from an animal of the control group (a), from the injection site of saline (b), and from the injection site of pVAXrbd at a dose of 100 μg (c).

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3. Fig. 2. Scheme of immunization of BALB/c mice with DNA vaccine pVAXrbd. Designations (here and below): IM – intramuscular, JI – jet injector, VNA – serum virus-neutralizing activity assay.

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4. Fig. 3. Analysis of the immune response in mice immunized with pVAXrbd. a – RBD-specific antibody (AB) titers based on ELISA results (see Experimental Section). b – Neutralizing activity of sera was determined on Vero E6 cells infected with the hCoV-19/Australia/VIC01/2020 strain by reducing the CPE of the virus. c – The number of IFN-γ-producing splenocytes was counted by ELISpot. d – Photographs of ELISpot wells: top row – splenocytes without peptide stimulation, bottom row – stimulated with a peptide mixture or mitogen. d – Quantitative determination of SARS-CoV-2 RNA in 10% lung homogenate of BALB/c mice on day 4 after infection. The results are presented as cycle threshold (Ct) values of real-time PCR. All graphs show median values with interval. Significance was calculated using nonparametric one-way Kruskal-Wallis analysis of variance; *p < 0.05, **p < 0.01.

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5. Fig. 4. Scheme of immunization of BALB/c mice with different doses of the pVAXrbd DNA vaccine.

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6. Fig. 5. Evaluation of adaptive immunity in mice immunized with different doses of the pVAXrbd plasmid (50, 100, and 200 μg) using a jet injector. a – RBD-specific antibody (AB) titers based on ELISA results. b – Neutralizing activity of sera was determined on Vero E6 cells infected with the hCoV-19/Australia/VIC01/2020 strain by reducing the CPE of the virus. c – Analysis of the neutralizing activity of mouse sera on pseudotyped viruses carrying the hCoV-19/Australia/VIC01/2020 S-glycoprotein on the surface. NT50 – 50% neutralization titer. d – Quantitative determination of SARS-CoV-2 RNA in the lungs of BALB/c mice on day 4 after infection. The results are presented as cycle threshold (Ct) values of real-time PCR. All graphs show median values with interval. Significance was calculated using nonparametric one-way Kruskal–Wallis analysis of variance; *p < 0.05, **p < 0.01, ***p < 0.001.

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