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Palyi, B.

Publications and source records attributed to Palyi, B..

6 recordsLinked to original sources

Nipah virus Malaysia and Bangladesh strain-induced pathogenesis in mice lacking type I interferon receptor signaling

Nipah virus (NiV) is a zoonotic highly pathogenic Paramyxovirus inducing lethal outbreaks of encephalitis and severe acute respiratory syndrome (SARS) with an average case-fatality rate of 75%. Two viral strains, NiV-Malaysia (NiV-Mal) and NiV-Bangladesh (NiV-Ban), associated to distinct geographical distribution, route of transmission, symptoms and lethality have been described. Due to the permanent threat of these emerging infections and the lack of approved therapeutics, it is crucial to improve our understanding regarding NiV-associated pathogenesis. Mice represent a small and accessible animal model, provided with numerous biological tools for the functional assessment of different genes related to antiviral response. Here, we analyze the susceptibility of mice deficient for type I interferon receptor (IFNAR KO) to infection with either NiV-Mal or NiV-Ban through intraperitoneal or intranasal routes. Our results show that IFNAR KO mice are susceptible to NiV-Ban infection via intraperitoneal route, although to a lesser extent than NiV-Mal, and develop encephalitis and pulmonary syndrome with viral propagation to different organs and lethal outcome in 60% of infected animals. In addition, intranasal administration of both viral strains led to a subclinical infection with viral replication in brain and lungs and production of virus-specific neutralizing antibodies. These results indicate IFNAR KO mice as a small animal model permitting comparative studies of the immunopathogenesis perpetrated by both NiV-Mal and NiV-Ban infections. Author summaryAvailability of small animal models represents a major issue to characterize virus-associated pathogenesis to further implement therapeutic strategies. Previous studies performed with NiV-Mal and NiV-Ban strains in wild-type (WT) mice did not indicate signs of NiV disease. However, when challenged with NiV-Mal, IFNAR KO mice suffered fatal outcomes, thus providing useful information on viral immunopathogenesis. Surprisingly, while being the more frequently re-emerging virus strain, no infection studies with NiV-Ban have been performed in IFNAR KO mice so far. Here, we sought comparing pathogenesis after NiV-Mal- and NiV-Ban-infection following IP and IN inoculations in IFNAR KO mice. Indeed, our results highlighted that contrary to fatal IP challenge, IN inoculation lead to a subclinical infection with both NiV strains. Moreover, we determined that NiV-Mal is more pathogenic than NiV-Ban following IP infection. Finally, distinct histopathological manifestations and tropism were associated with each viral strain and specific route of infection. Overall, our study implies that IFNAR KO mice represent a useful animal model to study strain-specific NiV pathophysiology.

microbiology↗

Barrier Function of the Vero E6 Cell Monolayer

Vero E6 cells are a standard model in virology research because of their broad susceptibility to viral infection. However, their ability to form functional barrier layers has received less attention. Here, we show that Vero E6 cells cultured on transwell inserts develop continuous monolayers with moderate transepithelial electrical resistance (TEER), reaching approximately 30 {Omega}{middle dot}cm2 in serum-containing medium and 45 {Omega}{middle dot}cm2 in virus production medium. The permeability coefficient (Papp) for 4 kDa FITC-dextran, a common measure for paracellular transport, is 3.7 x 10-6 cm/s, comparable to established endothelial models such as HUVEC or corneal endothelium. Immunofluorescence confirms the presence of tight junction (ZO-1) and adherens junction ({beta}-catenin) proteins. Chelation of calcium with EGTA causes a dose-dependent, reversible decrease in TEER and a marked increase in permeability, confirming the calcium sensitivity of the barrier. These findings suggest that Vero E6 cells serve as a valuable epithelial barrier model, facilitating studies on viral entry, immune evasion, and drug delivery.

cell biology↗

Purification and Immunogenicity of Nipah Virus-Like Particles from Insect Cells

Nipah virus (NiV) is an emerging high-fatality zoonotic threat lacking approved vaccines. Current virus-like particle (VLP) production methods rely on costly mammalian cell systems and non-scalable ultracentrifugation purification. We developed an efficient production system for enveloped NiVLPs by co-expressing structural proteins F, G, and M using a baculovirus expression vector system in Sf9 cells. A novel multi-step chromatographic purification process was established using monolith convective media, integrating steric exclusion chromatography with sequential cation and anion exchange steps. Purified NiVLPs were characterized by nanoparticle tracking analysis and transmission electron microscopy, then evaluated for immunogenicity in Syrian golden hamsters. The optimized process yielded enveloped particles of approximately 100-120 nm that morphologically resemble native NiV virions. A single 25 g NiVLP dose induced robust systemic anti-NiV G IgG responses within 14 days, demonstrating rapid immunogenicity suitable for outbreak response. However, neutralizing antibody titers against NiV remained limited compared to total IgG responses. This study establishes the first chromatography-based manufacturing platform for morphologically correct NiVLPs from insect cells. A deeper understanding of the immunity generated is needed to support their potential as a rapidly deployable vaccine platform against NiV.

bioengineering↗

Monitoring correlates of SARS-CoV-2 infection in cell culture using two-photon microscopy and a novel fluorescent calcium-sensitive dye

The organism-wide effects of viral infection SARS-CoV-2 are well studied, but little is known about the dynamics of how the infection spreads in time among or within cells due to the scarcity of suitable high-resolution experimental systems. Two-photon (2P) imaging combined with a proper subcellular staining technique has been an effective tool for studying mechanisms at such resolutions and organelle levels. Herein, we report the development of a novel calcium sensor molecule along with a 2P-technique for identifying imaging patterns associated with cellular correlates of infection damage within the cells. The method works as a cell viability assay and also provides valuable information on how the calcium level and intracellular distribution are perturbed by the virus. Moreover, it allows the quantitative analysis of infection dynamics. This novel approach facilitates the study of the infection progression and the quantification of the effects caused by viral variants and viral load.

microbiology↗

In vitro determination of inhibitory effects by humic substances complexing Zn and Se on SARS-CoV-2 virus replication

Humic substances are well known human nutritional supplement materials and play important performance-enhancing roles as animal feed additives, too. For decades, ingredients of humic substances have also been proven to carry potent antiviral effects against different viruses. Here, the antiviral activity of a humic substance containing ascorbic acid, Se- and Zn2+ ions intended as a nutritional supplement material was investigated against SARS-CoV-2 virus B1.1.7 Variant of Concern ("Alpha Variant") in a VeroE6 cell line. Results show that this combination has a significant in vitro antiviral effect at a very low concentration range of its intended active ingredients. Even picomolar concentration ranges of humic substances, vitamin C and Zn/Se ions in the given composition were enough to achieve fifty percent viral replication inhibition in the applied SARS-CoV-2 virus inhibition test.

microbiology↗

Topography, spike dynamics and nanomechanics of individual native SARS-CoV-2 virions

SARS-CoV-2, the virus responsible for the current COVID-19 pandemic, displays a corona-shaped layer of spikes which play fundamental role in the infection process. Recent structural data suggest that the spikes possess orientational freedom and the ribonucleoproteins segregate into basketlike structures. How these structural features regulate the dynamic and mechanical behavior of the native virion, however, remain unknown. By imaging and mechanically manipulating individual, native SARS-CoV-2 virions with atomic force microscopy, here we show that their surface displays a dynamic brush owing to the flexibility and rapid motion of the spikes. The virions are highly compliant and able to recover from drastic mechanical perturbations. Their global structure is remarkably temperature resistant, but the virion surface becomes progressively denuded of spikes upon thermal exposure. Thus, both the infectivity and thermal sensitivity of SARS-CoV-2 rely on the dynamics and the mechanics of the virus. One sentence summaryThe native coronavirus 2 displays a dynamic surface layer of spikes, a large mechanical compliance and unique self-healing capacity.

biophysics↗