DNA Vaccine Administered by Cationic Lipoplexes or by In Vivo Electroporation Induces Comparable Antibody Responses against SARS-CoV-2 in Mice

Peletta, Allegra; Prompetchara, Eakachai; Tharakhet, Kittipan; Kaewpang, Papatsara; Buranapraditkun, Supranee; Techawiwattanaboon, Teerasit; Jbilou, Tayeb; Krangvichian, Pratomporn; Sirivichayakul, Sunee; Manopwisedjaroen, Suwimon; Thitithanyanont, Arunee; Patarakul, Kanitha; Ruxrungtham, Kiat; Ketloy, Chutitorn; Borchard, Gerrit

doi:10.3390/vaccines9080874

Cited by 18 publications

(29 citation statements)

References 40 publications

(47 reference statements)

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“…A definite disadvantage of DNA vaccines is their low immunogenicity when injected as naked plasmid DNA [15,16]. A wide range of strategies have been tried to increase the immunogenicity of DNA vaccines, including packaging into liposomes; the use of "vaccine cocktails" containing the DNA vaccine as well as plasmids encoding adjuvant immunomodulatory proteins; and delivery by a gene gun, electroporation, or with a needle-free injector device [8,[17][18][19]. These methods help to solve the problem of immunogenicity, but there are sometimes safety problems, technological difficulties, and an increase in the cost of the developed vaccine.…”

Section: Introductionmentioning

confidence: 99%

Self-Assembled Particles Combining SARS-CoV-2 RBD Protein and RBD DNA Vaccine Induce Synergistic Enhancement of the Humoral Response in Mice

Borgoyakova

Karpenko

Рудометов

et al. 2022

IJMS

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Despite the fact that a range of vaccines against COVID-19 have already been created and are used for mass vaccination, the development of effective, safe, technological, and affordable vaccines continues. We have designed a vaccine that combines the recombinant protein and DNA vaccine approaches in a self-assembled particle. The receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 was conjugated to polyglucin:spermidine and mixed with DNA vaccine (pVAXrbd), which led to the formation of particles of combined coronavirus vaccine (CCV-RBD) that contain the DNA vaccine inside and RBD protein on the surface. CCV-RBD particles were characterized with gel filtration, electron microscopy, and biolayer interferometry. To investigate the immunogenicity of the combined vaccine and its components, mice were immunized with the DNA vaccine pVAXrbd or RBD protein as well as CCV-RBD particles. The highest antigen-specific IgG and neutralizing activity were induced by CCV-RBD, and the level of antibodies induced by DNA or RBD alone was significantly lower. The cellular immune response was detected only in the case of DNA or CCV-RBD vaccination. These results demonstrate that a combination of DNA vaccine and RBD protein in one construct synergistically increases the humoral response to RBD protein in mice.

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Section: Introductionmentioning

confidence: 99%

Self-Assembled Particles Combining SARS-CoV-2 RBD Protein and RBD DNA Vaccine Induce Synergistic Enhancement of the Humoral Response in Mice

Borgoyakova

Karpenko

Рудометов

et al. 2022

IJMS

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“…Liposomes were formulated using the TF method described elsewhere [ 9 , 14 ]. Briefly, lipids including DPPC, DOPE and DOTAP were dissolved in 4 mL chloroform:methanol (9:1) at a ratio of DPPC:DOPE:DOTAP (8:4:4) in a 50 mL round-bottom flask.…”

Section: Methodsmentioning

confidence: 99%

“…The particle size and Zeta-potential values were further examined. Liposome quantification was analyzed by the U-HPLC method as previously described [ 14 ].…”

Section: Methodsmentioning

confidence: 99%

“…This avoids the use of harsh processes such as sonication to reduce particle size and implement the homogeneity of the sample, permitting the encapsulation of fragile molecules such as nucleic acids [ 13 ]. Previously, a cationic liposomal formulation manufactured using thin-film rehydration and complexed with a SARS-CoV-2 spike (S) protein expressing plasmid DNA (pCMVkan-S) showed promising immunogenicity results in mice [ 14 ]. In lieu of upscale and transfer, a thin-film (TF) rehydration protocol was translated to a microfluidics (MF) method.…”

Section: Introductionmentioning

confidence: 99%

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Translating a Thin-Film Rehydration Method to Microfluidics for the Preparation of a SARS-CoV-2 DNA Vaccine: When Manufacturing Method Matters

et al. 2022

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Previous investigations conducted on a liposomal formulation for a SARS-CoV-2 DNA vaccine manufactured using the thin-film layer rehydration method showed promising immunogenicity results in mice. The adaptation of the liposomal formulation to a scalable and reproducible method of manufacture is necessary to continue the investigation of this vaccine candidate. Microfluidics manufacture shows high potential in method translation. The physicochemical characterization of the blank liposomes produced by thin-film layer rehydration or microfluidics were shown to be comparable. However, a difference in lipid nanostructure in the bilayer resulted in a significant difference in the hydration of the thin-film liposomes, ultimately altering their complexation behavior. A study on the complexation of liposomes with the DNA vaccine at various N/P ratios showed different sizes and Zeta-potential values between the two formulations. This difference in the complexation behavior resulted in distinct immunogenicity profiles in mice. The thin-film layer rehydration-manufactured liposomes induced a significantly higher response compared to the microfluidics-manufactured samples. The nanostructural analysis of the two samples revealed the critical importance of understanding the differences between the two formulations that resulted in the different immunogenicity in mice.

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“…Numerous companies and academic institutions across the globe have developed or are currently developing DNA/RNA vaccines for the SARS-CoV-2 spike protein ( 15 ). Generally, in the case of DNA vaccines, the full-length SARS-CoV-2 spike protein DNA sequence is inserted into a plasmid, and additional technologies, such as electroporation, can assist in making transfection more efficient ( 16 , 17 ). The spike protein DNA transfected into the human cell can then be transcribed and translated to create the trimeric spike protein, which, then, moves to the endoplasmic reticulum and Golgi apparatus for post-translational modification (e.g.…”

Section: Introductionmentioning

confidence: 99%

Are There Hidden Genes in DNA/RNA Vaccines?

et al. 2022

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Due to the fast global spreading of the Severe Acute Respiratory Syndrome Coronavirus – 2 (SARS-CoV-2), prevention and treatment options are direly needed in order to control infection-related morbidity, mortality, and economic losses. Although drug and inactivated and attenuated virus vaccine development can require significant amounts of time and resources, DNA and RNA vaccines offer a quick, simple, and cheap treatment alternative, even when produced on a large scale. The spike protein, which has been shown as the most antigenic SARS-CoV-2 protein, has been widely selected as the target of choice for DNA/RNA vaccines. Vaccination campaigns have reported high vaccination rates and protection, but numerous unintended effects, ranging from muscle pain to death, have led to concerns about the safety of RNA/DNA vaccines. In parallel to these studies, several open reading frames (ORFs) have been found to be overlapping SARS-CoV-2 accessory genes, two of which, ORF2b and ORF-Sh, overlap the spike protein sequence. Thus, the presence of these, and potentially other ORFs on SARS-CoV-2 DNA/RNA vaccines, could lead to the translation of undesired proteins during vaccination. Herein, we discuss the translation of overlapping genes in connection with DNA/RNA vaccines. Two mRNA vaccine spike protein sequences, which have been made publicly-available, were compared to the wild-type sequence in order to uncover possible differences in putative overlapping ORFs. Notably, the Moderna mRNA-1273 vaccine sequence is predicted to contain no frameshifted ORFs on the positive sense strand, which highlights the utility of codon optimization in DNA/RNA vaccine design to remove undesired overlapping ORFs. Since little information is available on ORF2b or ORF-Sh, we use structural bioinformatics techniques to investigate the structure-function relationship of these proteins. The presence of putative ORFs on DNA/RNA vaccine candidates implies that overlapping genes may contribute to the translation of smaller peptides, potentially leading to unintended clinical outcomes, and that the protein-coding potential of DNA/RNA vaccines should be rigorously examined prior to administration.

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DNA Vaccine Administered by Cationic Lipoplexes or by In Vivo Electroporation Induces Comparable Antibody Responses against SARS-CoV-2 in Mice

Cited by 18 publications

References 40 publications

Self-Assembled Particles Combining SARS-CoV-2 RBD Protein and RBD DNA Vaccine Induce Synergistic Enhancement of the Humoral Response in Mice

Self-Assembled Particles Combining SARS-CoV-2 RBD Protein and RBD DNA Vaccine Induce Synergistic Enhancement of the Humoral Response in Mice

Translating a Thin-Film Rehydration Method to Microfluidics for the Preparation of a SARS-CoV-2 DNA Vaccine: When Manufacturing Method Matters

Are There Hidden Genes in DNA/RNA Vaccines?

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