Project description:Zoonotic viruses are an omnipresent threat to global health. Influenza A virus (IAV) transmits between birds, livestock, and humans. Proviral host factors involved in the cross-species interface are well known, less is known about antiviral mechanisms that suppress IAV zoonoses. We observed CpG dinucleotide depletion in human IAV relative to avian IAV. Notably, human ZAP selectively depletes CpG-enriched viral RNAs with its cofactor KHNYN. ZAP is conserved in tetrapods but we uncovered that avian species lack KHNYN. We found that chicken ZAP does not affect IAV (PR8) or CpG enriched IAV. Human ZAP or KHNYN independently restricted CpG enriched IAV by overexpression in chicken cells or knockout in human cells. Additionally, mammalian ZAP-L and KHNYN also independently restricted an avian retrovirus (ROSV). Curiously, platypus KHNYN, the most divergent from eutherian mammals, was also capable of direct restriction of multiple diverse viruses. We suggest that mammalian KHNYN may be a bona fide restriction factor with cell-autonomous activity. Furthermore, we speculate that through repeated contact between avian viruses and mammalian hosts, protein changes may accompany CpG-biased mutations or reassortment to evade mammalian ZAP and KHNYN.
Project description:Zoonoses pose substantial global health risks, highlighting the need to better understand animal-to-human transmission. Reptiles are increasingly recognized as hosts of diverse pathogens, including many viruses. Despite this, reptile pathogens remain poorly understood in terms of their diversity, prevalence, and potential risk to humans. Yet human–reptile contact is increasing, driven in part by the global wildlife trade and pet industry. This underscores the need to better understand reptile-associated pathogens; however, models to study reptile viruses remain scarce. Here, we establish and characterize airway organoids derived from Python regius, providing an in vitro model of the reptile airway. Through de novo assembly of a Python regius reference genome, we characterized airway organoids at single-cell resolution, revealing diverse cell populations including ionocytes, ciliated, goblet, club, endocrine, tuft, and basal cells. The organoids supported productive infection with ball python nidovirus (BPNV) and mounted a robust epithelial antiviral response through induction of interferon-stimulated genes, cytokines and genes involved in chemical defense. As a proof-of-concept, antiviral drug treatment reduced BPNV levels, highlighting the model's utility for drug testing. By providing a reductionist system of the serpentine airway, these organoids constitute a physiologically relevant in vitro model to study reptile viruses and host–pathogen interactions in the native host. This system has potential veterinary applications, ecology of serpentes, and zoonotic disease research.
Project description:Zoonoses pose substantial global health risks, highlighting the need to better understand animal-to-human transmission. Reptiles are increasingly recognized as hosts of diverse pathogens, including many viruses. Despite this, reptile pathogens remain poorly understood in terms of their diversity, prevalence, and potential risk to humans. Yet human–reptile contact is increasing, driven in part by the global wildlife trade and pet industry. This underscores the need to better understand reptile-associated pathogens; however, models to study reptile viruses remain scarce. Here, we establish and characterize airway organoids derived from Python regius, providing an in vitro model of the reptile airway. Through de novo assembly of a Python regius reference genome, we characterized airway organoids at single-cell resolution, revealing diverse cell populations including ionocytes, ciliated, goblet, club, endocrine, tuft, and basal cells. The organoids supported productive infection with ball python nidovirus (BPNV) and mounted a robust epithelial antiviral response through induction of interferon-stimulated genes, cytokines and genes involved in chemical defense. As a proof-of-concept, antiviral drug treatment reduced BPNV levels, highlighting the model's utility for drug testing. By providing a reductionist system of the serpentine airway, these organoids constitute a physiologically relevant in vitro model to study reptile viruses and host–pathogen interactions in the native host. This system has potential veterinary applications, ecology of serpentes, and zoonotic disease research.
Project description:Background. Leptospirosis is among the most widespread zoonoses worldwide. Severe pulmonary hemorrhagic syndrome (SPHS) represents a serious complication of leptospirosis, with poor understanding of its underlying mechanisms and an urgent need for identification of effective biomarkers. Methods. A nested case-control analysis of the blood specimens obtained from two previous multi-center cohorts was conducted. Candidate microRNAs were initially discovered through a global profiling of 800 serum microRNAs, then validated using real-time polymerase-chain reactions. We further conducted a multi-omics analysis incorporating transcriptomic and proteomic data to identify enriched pathways through which the newly identified microRNAs could regulate. Findings. A total of 28 SPHS and 140 non-SPHS patients were evaluated by serum microRNA profiling, revealing distinct expression patterns between the two phenotypes. From the top 81 significantly expressed microRNAs, seven were selected for validation. Among these, miR-5010-3p and miR-147b-3p had area under the curve (AUC) values of 0.72 (95% CI: 0·62–0·81) and 0·66 (95% CI: 0·55–0·76) for discriminating SPHS. Notably, the two microRNAs could detect SPHS in patients who were yet to manifest chest radiograph shadows at the time of sample collection, and in a subgroup of patients who were recruited on day 2 of illness or earlier, with consistent AUC values. Integrated gene-protein pathway enrichment analysis revealed numerous pathways involving host immune responses. Among these, the tumor necrosis factor signaling pathway was the most significant with many of its member genes being targeted by miR-5010-3p or miR-147b-3p.
Project description:Novel coronavirus causing Covid-19 identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused pandemic in 2020. Although the virus and disease in humans has been thoroughly researched, so far there has not been animal model comparable to humans – genetically diverse species able to get infected and sick from Covid-19. The white-footed deermouse Peromyscus leucopus is a long-lived rodent and a key reservoir in North America for agents of several zoonoses including Lyme disease, babesiosis, anaplasmosis, and viral encephalitis. While persistently infected, this deermouse avoids apparent disability or diminished fitness. Its tolerance to infection with sometimes more than one pathogen makes P. leucopus comparable to bats. This study uses P. leucopus, LL colony stock, as a genetically diverse animal model for viral infection with SARS-CoV-2. We infected P. leucopus with SARS-CoV-2, collected plasma, lungs, and brain 3 and 6 days post-infection, and compared to control animals. P. leucopus mount an immune response against viral pathogens through production of neutralizing antibodies and genome-wide transcription of type I interferon stimulated genes in lungs compared to naïve animals. Viral RNA detection correlates with gene expression of type I interferon stimulated genes in response to viral infection in the brain. We report that diversity of outbred animals, their sex and age is reflected in the range of responses. These results show that P. leucopus is a viable animal model for SARS-CoV-2, particularly in research of viral infection of the brain.
Project description:Novel coronavirus causing Covid-19 identified as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused pandemic in 2020. Although the virus and disease in humans has been thoroughly researched, so far there has not been animal model comparable to humans – genetically diverse species able to get infected and sick from Covid-19. The white-footed deermouse Peromyscus leucopus is a long-lived rodent and a key reservoir in North America for agents of several zoonoses including Lyme disease, babesiosis, anaplasmosis, and viral encephalitis. While persistently infected, this deermouse avoids apparent disability or diminished fitness. Its tolerance to infection with sometimes more than one pathogen makes P. leucopus comparable to bats. This study uses P. leucopus, LL colony stock, as a genetically diverse animal model for viral infection with SARS-CoV-2. We infected P. leucopus with SARS-CoV-2, collected plasma, lungs, and brain 3 and 6 days post-infection, and compared to control animals. P. leucopus mount an immune response against viral pathogens through production of neutralizing antibodies and genome-wide transcription of type I interferon stimulated genes in lungs compared to naïve animals. Viral RNA detection correlates with gene expression of type I interferon stimulated genes in response to viral infection in the brain. We report that diversity of outbred animals, their sex and age is reflected in the range of responses. These results show that P. leucopus is a viable animal model for SARS-CoV-2, particularly in research of viral infection of the brain.