Acta virologica最新文献

Noroviruses (NoV) are enteric caliciviruses that have been detected in multiple species of mammals, including humans. Establishing an efficient in vitro cell culture system for human norovirus (HuNoV) remains a challenge; however, its replication has been reported in 3D cultured Caco-2 cells and a clone of Caco-2 cells (C2BBe1), human enteroids and human B cells. Isolated mouse intestinal villi, with large diversity of intestinal epithelial cells, are a primary cellular model that has been shown to be permissive for the infection and replication of enteric viruses such as rotaviruses. We hypothesized that they could allow the infection and replication of the human noroviruses. In this report, we indicate that the isolated villi model of the mouse intestine is effective for the infection study and replication of the human noroviruses from faeces and environmental matrices (water, vegetables and air). For successful infection, the virus needs to be activated with trypsin. The virus has an average replicative cycle of 10 h, although viral particles with infectious capacity are found already at 2 hours post infection (2 h.p.i.). The model is efficient in obtaining abundant biological material and is ideal for studying the biological activity of the human noroviruses in the same cell model or for generating antibodies. Keywords: human norovirus; intestinal epithelial cells; intestinal villus isolation; norovirus isolated from water; norovirus isolated from plants; norovirus isolated from air.

High-throughput RNA sequencing (RNA-seq) analysis of samples from Mallotus japonicus, a traditional medicinal plant, yielded two novel RNA viruses tentatively named Mallotus japonicus virus A (MjVA) and Mallotus japonicus virus B (MjVB). The MjVA and MjVB genomes encode proteins showing amino acid sequence similarities to those of poleroviruses (the genus Polerovirus, the family Solemoviridae) and amalgaviruses (the genus Amalgavirus, the family Amalgaviridae), respectively. The MjVA genome contains seven highly overlapping open reading frames, which are translated to seven proteins through various translational mechanisms, including -1 programmed ribosomal frameshifting (PRF) at the slippery motif GGGAAAC, non-AUG translational initiation, and stop codon readthrough. The MjVB genome encodes two proteins; one of which is translated by +1 PRF mechanism at the slippery motif UUUCGN. The abundance analysis of virus-derived RNA fragments revealed that MjVA is highly concentrated in plant parts with well-developed phloem tissues as previously demonstrated in other poleroviruses, which are transmitted by phloem feeders, such as aphids. MjVB, an amalgavirus generally transmitted by seeds, is distributed in all samples at low concentrations. Thus, this study demonstrates the effectiveness and usefulness of RNA-seq analysis of plant samples for the identification of novel RNA viruses and analysis of their tissue distribution. Keywords: Polerovirus; Amalgavirus; Mallotus japonicus; RNA virus; viral genome; programmed ribosomal frameshifting.

The major protective immune response against viruses is the production of type I and III interferons (IFNs). IFNs induce the expression of hundreds of IFN-stimulated genes (ISGs) that block viral replication and further viral spread. In this report, we analyzed the expression of IFNs and some ISGs (MxA, PKR, OAS-1, IFIT-1, RIG-1, MDA5, SOCS-1) in alveolar epithelial cells (A549) in response to infection with influenza A viruses (A/California/07/09 (H1N1pdm); A/Texas/50/12 (H3N2)); influenza B virus (B/Phuket/3073/13); adenovirus type 5 and 6; or respiratory syncytial virus (strain A2). Influenza B virus had the ability to most rapidly induce IFNs and ISGs as well as to stimulate excessive IFN-α, IFN-β and IFN-λ secretion. It seems curious that IAV H1N1pdm did not induce IFN-λ secretion, but enhanced type I IFN and interleukin (IL)-6 production. We emphasized the importance of the negative regulation of virus-triggered signaling and cellular IFN response. We showed a decrease in IFNLR1 mRNA in the case of IBV infection. The attenuation of SOCS-1 expression in IAV H1N1pdm can be considered as the inability of the system to restore the immune status. Presumably, the lack of negative feedback loop regulation of proinflammatory immune response may be a factor contributing to the particular pathogenicity of several strains of influenza. Keywords: lambda interferons; MxA; influenza; respiratory syncytial virus; A549 cells.

Equine herpesvirus 1 (EHV1) infection is a global health problem in equines and the virus is responsible for abortions, respiratory disease and myeloencephalitis in horses. Disease management requires proper biosecurity and immunoprophylactic measures. Vaccines strengthening both arms of immunity are essential for proper control and there has been a continuous focus in this area for generation of better vaccines. Here we report construction of bacterial artificial chromosome (BAC) clone of EHV-1 strain Tohana for mutagenesis of the virus and generation of gE gene deletion mutant EHV1. The BAC clone was generated by inserting the mini-F plasmid replacing ORF71 of EHV1 and transforming into E. coli for generation of EHV1-BAC. The infectious virus was regenerated from EHV-1 BAC DNA in RK13 cells. To check utility of EHV1-BAC, we have generated mutant EHV1 by deleting the virulence-associated gE gene. The mutant virus (vToHΔgE) showed significantly reduced plaque size without affecting replication efficiency. Pathological evaluation of lesions in BALB/c mice infected with vToHΔgE revealed reduction in clinical signs and pathology in comparison to the wild-type virus. Generation of infectious BAC of EHV1 and its usage in construction of attenuated viruses shows potential of the technology for development of indigenous modified live vaccine for EHV1. Keywords: quine herpesvirus 1; bacterial artificial chromosome (BAC); mutation; glycoprotein E; vaccine.

Parvoviruses affect both vertebrates and invertebrates, and can be both detrimental and benign to the host. Numerous studies about parvovirus-induced apoptotic cell death have been researched and reported. In most parvovirus infections, cell death heightens the virus dissemination and causes tissue damage, often leading to disease. Cell cycle arrest also induces cytopathic effects in infected cells and is sometimes a prerequisite to apoptotic cell death. Cell death mechanisms caused by parvovirus infections vary depending on the infecting parvovirus strain and the cell lines involved. Apo-ptosis, however, is a frequent form of cell death induced by parvoviruses. The non-structural protein 1 (NS1) is a major contributor to parvovirus infection-induced cell death. However, other proteins such as the 11 kDa, NP1 and viral genome replication can also induce cell death. Understanding the mechanisms involved in parvovirus cell death, and host response is important in the development of treatment for cytopathic parvoviruses. This review article discusses parvovirus-induced apoptotic cell death and the mechanisms involved. Keywords: apoptosis; cell cycle arrest; cell death; parvovirus; viral protein.

Sweepoviruses represent a phylogenetic group of begomoviruses that cause significant sweet potato (Ipomoea batatas) production losses in various countries across the world. For rapid identification of sweepoviruses, we developed a technique based on isothermal recombinase polymerase amplification in conjunction with lateral flow dipsticks (RPA-LFD). The optimum reaction conditions for the RPA were 20 min incubation at 37°C. The RPA-LFD specifically detected distinct sweepovirus species, with no other viruses infecting sweet potato causing a cross-reaction. The detection limit of the RPA-LFD was 1.0×104 copies of the target DNA molecule per reaction, and it exhibited a 10-fold greater sensitivity than the conventional PCR. Furthermore, when coupled with an alkaline polyethylene glycol-based crude genomic DNA extraction, the entire procedure was completed in 30 min without the use of any special instruments other than a water bath. Therefore, the RPA-LFD technique is a potential sweepovirus diagnostic tool that can be used in the field with fewer available resources. Keywords: detection; sweepoviruses; recombinase polymerase amplification; lateral flow dipstick.

 Newcastle disease virus (NDV) is a paramyxovirus that is highly pathogenic to poultry causing severe economic loss worldwide. The non-structural V protein is one of the virulence factors of the virus. It antagonises the interferon of the host innate immunity in order to allow successful virus replication in the host cells. However, detailed investigation of recombinant NDV expressing mutated V protein is scarce. In this study, a mesogenic recombinant NDV expressing GFP (rAF-GFP) was used to investigate the relation of V protein mutation on virus pathogenicity. Site-directed mutagenesis was performed using overlapping PCR to introduce four premature stop codons 456G>T, 537G>T, 624C>T and 642G>T in the V gene reading frame. The virus was then rescued and propagated in embryonated chicken eggs. However, instead of the substituted thymine, this nucleotide was mutated into cytosine in three rescued mutants, while 537G>T mutant could not be rescued. As a result, the premature stop codon was substituted with other amino acid and the V protein was expressed in full length. The pathogenicity type of the rAF (456G>T>C), rAF (624C>T>C), and rAF (642G>T>C) mutants remained to be as in mesogenic strains, suggesting that substituted amino acids were functionally interchangeable with the original amino acids present in V protein. It appears that an intact V protein is important for the virus survival. This study explored the possibility of V protein mutation in NDV through exploiting genetic engineering and warrants a further investigation on modifying mutations on a conserved protein in NDV or other paramyxoviruses. Keywords: Paramyxoviridae; Newcastle disease virus; V protein; C terminal; virulence factor.

Slovakia is a country with only 5.45 million inhabitants. However, the past two years of the COVID-19 pandemic have shown huge inter-regional differences. These were represented by different numbers of diagnosed SARS-CoV-2 cases and the vaccination rates in the regions, as well as by the willingness of the inhabitants to comply with anti-pandemic measures or to undergo testing. The occurrence of such regional disparities provided a rational basis for monitoring the epidemic situation within smaller areas, e.g. at city level. Trencin is a medium-sized Slovak county town with about 55 000 inhabitants. The city administration gave its residents the opportunity to assess their current level of antibodies against the SARS-CoV-2 virus, and received an additional benefit in the form of data on the real epidemic situation in the city, which helped in further management of anti-pandemic measures. The primary aim of the study, conducted in January and February 2022, was to determine the levels of antibodies against the SARS-CoV-2 virus in the inhabitants of Trencin. The results showed that 75% of the study participants, representing the adult population of the city, had detectable IgG antibodies against the SARS-CoV-2 spike protein. Noteworthy, at the time of the study, 13% of the Trencin city population who were unaware of overcoming COVID-19 had specific antibodies against the virus. Furthermore, the antibody levels in recovered unvaccinated subjects increased not only with the severity of their COVID-19 symptoms, but also after multiple recoveries from the disease. On the other hand, the severity of side effects after vaccination did not influence the antibody levels. The results of the study are in line with the current view that hybrid immunity (vaccination plus SARS-CoV-2 infection in any order) offers greater protection than immunity elicited by vaccination or COVID-19 separately. Keywords: SARS-CoV-2 coronavirus; COVID-19; ELISA; seroprevalence; antibodies; vaccination.