Biology subjects

Petrovska, R.

Publications and source records attributed to Petrovska, R..

3 recordsLinked to original sources

bioRxiv2026

A plant virus-based virus like-particle platform for therapeutic vaccination targeting IL-23A

Interleukin-23 (IL-23) is a pivotal cytokine in the pathogenesis of immune-mediated inflammatory diseases, making it an attractive therapeutic target. Vaccination against self-antigens offers an alternative to monoclonal antibody (mAb) therapies by eliciting endogenous Ab responses. Here, we report the development of a virus-like particle (VLP)-based vaccine targeting murine IL-23A (mIL23A) using the eggplant mosaic virus (EMV) coat protein as a carrier. mIL23A was incorporated either as a direct fusion or within a mosaic system, with the latter supporting efficient VLP assembly. Alternative antigen presentation strategies employed synthetic coiled-coil and zipper pairs. All constructs were expressed in E. coli, purified, and characterized by SDS-PAGE, TEM, and DLS. Immunization of BALB/c mice elicited robust mIL23A-specific Abs, with mosaic VLPs inducing high-avidity, Th1-biased responses. Encapsidated RNA within VLPs was double-stranded and stimulated TLR3, enhancing immunogenicity. Scale-up in bioreactors preserved VLP integrity and antigen incorporation, yielding 1.72 mg per gram of cells. These results establish EMV VLPs as a flexible and scalable platform for presenting challenging antigens and provide a promising approach for therapeutic vaccination against self-antigens.

immunology↗

bioRxiv2022

Bacteriophage-derived dsRNA exerts anti-SARS-CoV-2 activity in vitro and in Golden Syrian hamsters in vivo

PurposeBacteriophage-derived dsRNA, also known as Larifan, is nationally well-known broad-spectrum antiviral medication. The goal of this study was to ascertain the antiviral activity of Larifan against the novel SARS-CoV-2. MethodsThe antiviral activity of Larifan against SARS-CoV-2 in vitro was measured in human lung adenocarcinoma (Calu3) and primary human small airway epithelial cells (HSAEC) using cytopathic effect assay, viral RNA copy number detection by digital droplet PCR (ddPCR) and infectious virus titration in cells supernatants in Vero E6 cells by end-point titration method. The antiviral effect of Larifan in vivo was detected in SARS-CoV-2 infection model in Golden Syrian hamsters. Larifan (5 mg/kg) was administered either subcutaneously or intranasally twice before and after virus infection with a 24-hour interval between doses. The viral RNA copies and infectious virus titre were detected in animal lungs at day three and five post-infection using ddPCR and end-point titration in Vero E6 cells, respectively. Histopathology of lungs was analysed as well. ResultsLarifan inhibited SARS-CoV-2 replication in Calu3 cells both after the drug addition pre- and post-infection with a substantial drop in the supernatant viral RNA copy numbers from eight (p = 0.0013) to twenty (p = 0.0042) times, respectively. Similarly, infectious virus titre in Vero E6 cells dropped by 3.6log10 TCID50 and 2.8log10 TCID50 after the drug addition pre- and post-infection, respectively. In HSAEC, Larifan inhibited SARS-CoV-2 replication at the similar level. Larifan also markedly reduced virus numbers in the lungs of infected hamsters (p = 0.0032) both at day three and five post-infection with a more pronounced effect after intranasal administration reaching a drop by 2.7log10 at day three and 2.0log10 at day five. The administration of Larifan also reduced the amount of infections virus titer in lungs (p = 0.0039) by 4.3log10 TCID50 and 2.8log10 TCID50 at day three and five post-infection, respectively. Improvements in the infection-induced pathological lesion severity in the lungs of animals treated with Larifan were also demonstrated by histological analyses. ConclusionsThe inhibition of SARS-CoV-2 replication in vitro and the reduction of the viral load in the lungs of infected hamsters treated with Larifan alongside the improved lung histopathology, suggests a potential use of Larifan in controlling the COVID-19 disease in humans.

microbiology↗

bioRxiv2020

Metformin strongly affects gut microbiome composition in high-fat diet-induced type 2 diabetes mouse model of both sexes

Effects of metformin, the first-line drug for type 2 diabetes therapy, on gut microbiome composition in type 2 diabetes have been described in various studies both in human subjects and animals. However, the details of the molecular mechanisms of metformin action have not been fully understood. Moreover, there is a significant lack of information on how metformin affects gut microbiome composition in female mice models, as most of the existing studies have focused on males only. Our study aimed to examine metformin-induced alterations in gut microbiome diversity and composition of high-fat diet-induced type 2 diabetes mouse model, employing a randomized block, factorial study design, and including 24 experimental units allocated to 8 treatment groups. We performed shotgun metagenomic sequencing using DNA obtained from fecal samples representing gut microbiome before and after ten weeks-long metformin treatment. We identified 100 metformin-related differentially abundant species in high-fat diet-fed mice before and after the treatment, with most of the species abundances increased. In contrast, no significant changes were observed in control diet-fed mice.We also observed sex-specific differences in response to metformin treatment. Males experienced more pronounced changes in metabolic markers, while, in females, the extent of changes in gut microbiome representatives was more marked, indicated by 53 differentially abundant species with more remarkable Log fold changes compared to the combined-sex analysis. Our results suggest that both sexes of animals should be included in future studies focusing on metformin effects on the gut microbiome.

microbiology↗