Abstract
Contemporary SARS-Cov-2 pandemic, besides its dramatic global influence on the human race including health care systems, economies, and political decisions, opened a window for the global experiment with human vaccination employing novel injectable vaccines providing predominantly specific IgG response with little knowledge of their impact on the mucosal immunity. However, it is widely accepted that protection against the pathogens at the gates of the infection - on mucosal surfaces—predominantly rely on an IgA response. Some genetically modified bacteria, including probiotics, represent attractive vehicles for oral or nasal mucosal delivery of therapeutic molecules. Probiotic-based vaccines for mucous membranes are easy to produce in large quantities; they have low cost, provide quite a long T-cell memory, and gut IgA response to oral vaccines is highly synchronized and strongly oligoclonal. Here we present a study demonstrating construction of the novel SARS-Cov-2 vaccine candidate employing the gene fragment of S1 SARS-Cov-2 gene. This DNA fragment was inserted in frame into major pili protein gene with d2 domain of enterococcal operon encoding for pili. The DNA sequencing proved the presence of the insert in enterococcal genome. RNA transcription, immunoprecipitation, and immune electron microscopy with human sera obtained from the SARS-Cov-2 patients demonstrated expression of SARS-Cov-2 antigens in bacteria. Taken together the data obtained allowed considering this genetically modified probiotic strain as an interesting candidate for vaccine against SARS-Cov-2.
Highlights
The onset of the SARS-Cov-2 pandemic required the urgent preventive measures to limit the spread of the virus has accelerated the development of antiviral vaccines
The DNA sequence corresponding to this S protein area was synthesized and used to make the expression plasmid pQE-sarsS and suicidal plasmid pentF-sarsS which was constructed for insertion into the bacterial genome (Supplementary Figure S1)
To confirm the expression of the inserted sarsS gene fragment in bacterial DNA, we studied the expression of mRNA using real-time reverse transcriptase polymerase chain reaction (PCR) with SarsS specific primers K1 j K2. (Table 1)
Summary
The onset of the SARS-Cov-2 pandemic required the urgent preventive measures to limit the spread of the virus has accelerated the development of antiviral vaccines. The majority of the vaccines on the Probiotic Vaccine Against SARS-Cov-2 market indifferently on vaccine making approach, rely on the needle injection of the vaccine hoping for the immune recognition of the viral antigens and for establishment of the cellular and adaptive immune responses to the pathogen in case of its appearance in the organism Such approaches produce a weak immune response on the mucous membranes at the gate of the infection which is oral or gut mucosa allowing the virus to enter the organism. This makes it possible to spread the disease through a fully vaccinated population and provide the possibility of artificial induction of the appearance of viral variants under the pressure of the targeted immune response inflicted by the vaccination. We describe the construction and preliminary study of novel mucosal vaccine candidate with enterococcal probiotic as the vector for viral antigens providing immune recognition of SARS-Cov-2
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