Keskin, S. | bioRxiv Science

bioRxiv2025

Complete Protection Against Lethal Crimean-Congo Hemorrhagic Fever Virus (CCHFV) Challenge in Mice Afforded by Immunization with Modified mRNA Encoding the Nucleocapsid Protein (NP) Alone

BackgroundThe Crimean-Congo Hemorrhagic Fever Virus (Orthonairovirus haemorrhagiae) causes a hemorrhagic fever with mortality rates reaching up to 40%. For years, this virus has maintained its position among the top priority pathogens identified by the World Health Organization (WHO). This is due to its endemic presence across a vast region--from Africa and Spain to the Balkans, the Middle East, and throughout Asia--its potential for human-to-human transmission, and the lack of an effective and approved vaccine or treatment. Therefore, the development of an effective vaccine against CCHFV is of critical importance. Building on the success of mRNA-based vaccines during the Coronavirus Disease 2019 (COVID-19) pandemic, this study reports the development of a messenger ribonucleic acid (mRNA) vaccine candidate expressing the nucleocapsid protein (NP) of CCHFV. The CCHFV NP in vitro transcript (IVT) was designed with pseudouridine ({Psi}) nucleoside modification. MethodsAs part of the preclinical characterization of the IVT vaccine candidate, the biochemical and immunological properties of NP were confirmed in Huh-7 cells transfected with IVT NP-{Psi}mRNA. Afterwards, the efficacy of IVT NP-{Psi}mRNA immunization was evaluated in immunocompetent BALB/c and transiently immunosuppressed (IS) C57BL/6 mice. In CCHFV challenge studies, IS C57BL/6 mice were used. IS C57BL/6 mice were immunized intramuscularly with 2 doses of NP-{Psi}mRNA, either naked or encapsulated in Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, administered 14 days apart. FindingsHigh levels of CCHFV NP-specific humoral (IgM and IgG) and cellular (cytokine and lymphoproliferative) responses were demonstrated in BALB/c mice immunized with IVT NP-{Psi}mRNA. In challenge experiments, 100% protection was achieved by both the naked and PLGA-encapsulated IVT NP-{Psi}mRNA immunizations. Additionally, full protection was observed in mice immunized with inactivated CCHFV, whereas only 20% protection was detected in the unmodified IVT NP-mRNA vaccinated animals. In the protected mice, viral clearance was observed in the spleen, liver tissues, and blood on day 14 post-challenge. InterpretationThis study demonstrates that NP, the most abundant protein of the virus, is capable of providing full protection as a standalone vaccine candidate. Furthermore, our report represents a crucial milestone in identifying a future vaccine candidate and paves the way for subsequent clinical studies. FundingsThis study was supported by the Bezmialem Vakif University Scientific Research Project BAP#20220402. Additionally, this study received support from TUBITAK 2211/C. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=115 SRC="FIGDIR/small/649815v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@6e5aa4org.highwire.dtl.DTLVardef@100f150org.highwire.dtl.DTLVardef@e2f004org.highwire.dtl.DTLVardef@fbcf7b_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2024

Promoting Longevity in Aged Liver through NLRP3 Inflammasome Inhibition Using Tauroursodeoxycholic Acid (TUDCA) and SCD Probiotics

This study investigates the combined impact of SCD Probiotics and tauroursodeoxycholic acid (TUDCA) on the biomolecular makeup, histological changes and levels of inflammasome in the liver tissue of 24-month-old male Sprague-Dawley rats. By administering TUDCA (300 mg/kg, intravenously) and SCD Probiotics (3 mL (1 x 108 CFU), orally) daily for a week, the researchers employed ATR-FTIR spectroscopy along with machine learning approaches such as Linear Discriminant Analysis (LDA) and Support Vector Machine (SVM) to analyze the biomolecular profiles. In addition, the study measured the expression levels of inflammasome markers NLRP3, ASC, Caspase-1, IL18, and IL1{beta} using RT-qPCR and examined liver sections for histopathological changes and NLRP3 inflammasome activation. The results revealed significant differences in the levels of lipids, proteins, and nucleic acids, with TUDCA having a noteworthy impact on enhancing lipid bands and reducing cholesterol ester bands, while SCD Probiotics showed the opposite effects. Furthermore, TUDCA was found to decrease the acyl chain length of fatty acids and improve protein conformation, whereas SCD Probiotics increased both the acyl chain length and protein phosphorylation ratio, suggesting a decrease in lipid and protein dynamics from both treatments. The histological assessments showed significant reductions in cellular degeneration, lymphatic infiltration, hepatic fibrosis, and the immunoreactivity of NLRP3 and ASC in the treated groups. SCD Probiotics exhibited a marked reduction in inflammasome-related gene expressions, and the lowest gene expression levels were observed in the group receiving both treatments. Despite an increase in serum AST and LDH levels across all groups, only the SCD Probiotics group showed an increase in albumin levels. The findings suggest that SCD Probiotics, TUDCA, and their combined administration may provide a promising avenue for therapeutic interventions in age-associated liver conditions and may mitigate age-related liver fibrosis while enhancing liver functionality.

molecular biology↗