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A six-proline stabilized trimeric spike DNA vaccine induces robust immunity and protects against SARS-CoV-2 delta challenge in K18-hACE2 transgenic mice - 19/07/26

Doi : 10.1016/j.biopha.2026.119679 
Aytül Gül-Mete a, b, Ceren Gül b, c, Tuğba Karakavuk b, c, Muhammet Karakavuk b, d, e, Hüseyin Can b, e, f, Hivda Ülbeği-Polat g, Müge Serhatlı g, Hilal Yazıcı g, Şenay Köm g, Arzu Taş Ekiz g, İrem Abaci-Bayraktar h, Özge Aksoy i, Hakan Enül j, Cumhur Adıay j, Serdar Uzar j, Fahriye Saraç j, Aysu Değirmenci Döşkaya b, e, k, Adnan Yüksel Gürüz b, e, k, Mert Döşkaya b, e, k, , Elif Esin Hameş a, l
a Department of Bioengineering, Graduate School of Natural and Applied Sciences, Ege University, İzmir, Türkiye 
b Vaccine Development Application and Research Center, Ege University, İzmir, Türkiye 
c Department of Biotechnology, Graduate School of Natural and Applied Sciences, Ege University, İzmir, Türkiye 
d Ödemiş Vocational Training School, Ege University, İzmir, Türkiye 
e Department of Vaccine Studies, Institute of Health Sciences, Ege University, İzmir, Türkiye 
f Department of Biology, Molecular Biology Section, Faculty of Science, Ege University, İzmir, Türkiye 
g Life Science, Marmara Research Center, TÜBITAK, Kocaeli, Türkiye 
h Department of Biotechnology, Biotechnology Institute, Gebze Technical University, Kocaeli, Türkiye 
i Institute of Science, Department of Molecular Biology and Genetics, Yıldız Technical University, İstanbul, Türkiye 
j Pendik Veterinary Control Institute, İstanbul, Türkiye 
k Department of Parasitology, Faculty of Medicine, Ege University, İzmir, Türkiye 
l Department of Bioengineering, Faculty of Engineering, Ege University, İzmir, Türkiye 

Correspondence to: Vaccine Development Application and Research Center, Ege University, Bornova, İzmir 35100, Türkiye. Vaccine Development Application and Research Center, Ege University Bornova İzmir 35100 Türkiye

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Abstract

Despite the widespread deployment of vaccines, SARS-CoV-2 continues to pose a serious global health challenge due to the emergence of vaccine-resistant variants. DNA vaccines represent a critical platform for pandemic preparedness due to their ability to stimulate both arms of the immune response, their superior temperature stability, and their rapid scalability. In this study, we describe the immunogenicity and protective efficiency of a DNA vaccine encoding a six-proline stabilized trimeric spike protein (pSTDSpike) designed against the SARS-CoV-2 Delta variant. The pSTDSpike encodes Delta Spike protein stabilized in a prefusion trimeric conformation via six proline (6 P) and two alanine substitutions, complemented by a T4 fibritin foldon domain. Robust expression of the STDSpike protein was confirmed in vitro using HEK293T cells. To assess immunogenicity, BALB/c mice were administered three times via intramuscular (IM) injection or intradermal injection coupled with electroporation (ID+EP). Both vaccinations elicited potent humoral immune response, as evidenced by high anti-S1 IgG titers (∼22,000 for IM and ∼83,000 for ID+EP), >  80% inhibition activity, and significantly elevated neutralizing antibody titers (194.01 for IM and 337.79 for ID+EP). Furthermore, pSTDSpike induced a robust Th1-biased cellular response, characterized by significantly elevated IFN-γ secretion and increased cytotoxic CD8 + T-cells in restimulated splenocyte cultures. In a lethal challenge model using K18-hACE2 transgenic mice, pSTDSpike vaccination conferred 77.7% protection, without any detectable viral load, pneumonia or alveolar inflammation in surviving animals. These results suggest that the STDSpike DNA vaccine platform is highly effective and adaptable for addressing emerging variants and future pandemic threats.

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Graphical abstract




Il testo completo di questo articolo è disponibile in PDF.

Highlights

A Novel DNA vaccine encoding a six-proline stabilized trimeric Spike protein of SARS-CoV-2 Delta variant has been developed.
ID injection of DNA vaccine using electroporation-yielded higher IgG and neutralizing antibodies compared to IM injection.
Vaccination induced a robust cellular immune response characterized by elevated IFN- γ secretion and increased CD8 T-cells.
DNA vaccine conferred 77.7% protection in lethal challenge models using K18-hACE2 transgenic mice.
Mice challenged with SARS-CoV-2 Delta variant showed no detectable viral load or pulmonary inflammation.

Il testo completo di questo articolo è disponibile in PDF.

Keywords : DNA vaccine, SARS-CoV-2 Delta variant, Spike protein stabilization, Immunogenicity, Neutralization, K18-hACE2 mice


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