Increased neurovirulence of omicron BA.5 and XBB variants over BA.1 in K18-hACE2 mice and human brain organoids

Stewart, Romal; Ellis, Sevannah A.; Yan, Kexin; Dumenil, Troy; Tang, Bing; Nguyen, Wilson; Bishop, Cameron; Larcher, Thibaut; Parry, Rhys; Sullivan, Rodney N.; Lor, Mary; Khromykh, Alexander A.; Meunier, Frédéric A.; Rawle, Daniel J.; Suhrbier, Andreas

doi:10.1101/2022.12.22.521696

Cited by 5 publications

(6 citation statements)

References 182 publications

(440 reference statements)

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“…Infection of human cortical brain organoids hBOs were reprogrammed from adult dermal fibroblasts (HDFa, Gibco, C0135C) using the CytoTune-iPS 2.0 Sendai Reprogramming Kit (Invitrogen, A16518) 36 , and were grown using the CelVivo Clinostar incubator (Invitro Technologies) as described 37 . On the day of infection, ~30-day-old hBOs were transferred from each Clinoreactor into an ultra-low-binding 24-well plate (one hBO per well), and each hBO was infected with 10 5 CCID 50 of either JEV Nakayama , JEV FU , JEV NSW/22 , MVEV TC123130 , IMOJEV 38 , or YFV 17D 25 for ~4 h. For shorter-term culture (up to 4 days for viral growth kinetics), virus inoculum was removed, and hBOs were washed twice with media in the well, before 1 ml differentiation media was added to each well, and the 24-well plate was placed within a humidified tissue culture incubator at 37 °C, 5% CO 2 for up to 4 days.…”

Section: Histopathology and Immunohistochemistrymentioning

confidence: 99%

Characterisation of a Japanese Encephalitis virus genotype 4 isolate from the 2022 Australian outbreak

Nguyen,

Gyawali,

Stewart

et al. 2024

npj Viruses

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Human infections with the Japanese encephalitis virus (JEV) are a leading cause of viral encephalitis. An unprecedented outbreak of JEV genotype 4 was recently reported in Australia, with an isolate (JEVNSW/22) obtained from a stillborn piglet brain. Herein we conduct a thorough characterization of JEVNSW/22 in three different mouse strains and in human cortical brain organoids (hBOs), and determined the ability of JEVNSW/22 to be neutralized by sera from humans vaccinated with IMOJEV. JEVNSW/22 was less virulent than JEVFU (genotype 2) and JEVNakayama (genotype 3) in C57BL/6J mice and in interferon regulatory factor 7 deficient (Irf7−/−) mice, with infection of wild-type and knockout murine embryonic fibroblasts indicating JEVNSW/22 is more sensitive to type I interferon responses. Irf7−/− mice provide a new model for JEVNSW/22, showing higher viremia levels compared to C57BL/6J mice, and allowing for lethal neuroinvasive infection. All JEV strains were universally lethal in Ifnar−/− mice by day 3, with histological signs of brain hemorrhage, but no other lesions. There were no indications of brain infection in Ifnar−/− mice, with viral protein detected in blood vessels, but not neurons. All JEV isolates showed robust cytopathic infection of human cortical brain organoids, albeit lower for JEVNSW/22. IMOJEV vaccination in humans induced antibodies capable of neutralizing JEVNSW/22, although, for all JEV strains, cross-neutralization titers declined with increasing divergence from IMOJEV in the envelope amino acid sequences. Overall, our study establishes JEVNSW/22 mouse and hBO models of infection, allowing for possible lethal neuroinvasive infection in mice that was rarer than for other JEV genotypes. JEV vaccination regimens may afford protection against this newly emerged JEV genotype 4 strain, although neutralizing antibody responses are sub-optimal.

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Section: Histopathology and Immunohistochemistrymentioning

confidence: 99%

Characterisation of a Japanese Encephalitis virus genotype 4 isolate from the 2022 Australian outbreak

Nguyen,

Gyawali,

Stewart

et al. 2024

npj Viruses

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“…The omicron BA.5 isolate, SARS-CoV-2QIMR03 (SARS-CoV-2/human/AUS/QIMR03/2022) belongs to the BE.1 sublineage (GenBank: OP604184.1) and was obtained at QIMR Berghofer MRI from nasal swabs from a consented COVID-19 patient (Stewart et al, 2023). Virus stocks were propagated in Vero E6 cells, stocks and tissue culture supernatants were checked for endotoxin (Hirata et al, 2018;Johnson et al, 2005) and mycoplasma (MycoAlert, Lonza) (La Linn et al, 1995).…”

Section: The Sars-cov-2 Omicron Ba5 Virus Isolatementioning

confidence: 99%

“…Each group of mice within an experiment had a similar age range and distribution, with the mean age for each group not differing by more than 1 week. Mice were weighed and overt disease symptoms scored as described (Stewart et al, 2023). Mice were euthanized using CO2, and tissue titres determined using CCID50 assays and Vero E6 cells (Yan et al, 2021).…”

Section: Mace2-hace2 Mice and Infectionmentioning

confidence: 99%

“…The best described mouse model for SARS-CoV-2 infection is the K18-hACE2 model which expresses the SARS-CoV-2 receptor, human ACE2 (hACE2), from the keratin 18 promoter (K18). This model is generally lethal within several days, as it usually leads to brain infection and ensuing weight loss that reaches ethically defined criteria for euthanasia (Dumenil et al, 2023;Mills et al, 2021;Stewart et al, 2023). A non-lethal, less severe model is the mACE2-hACE2 model, wherein hACE2 is expressed from the mouse ACE2 promoter (mACE2) (Bao et al, 2020).…”

Section: The Mace2-hace2 Omicron Ba5 Mouse Model Of Sars-cov-2 Infect...mentioning

confidence: 99%

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Microplastics dysregulate innate immunity in the SARS-CoV-2 infected lung

Bishop,

Yan,

Nguyen

et al. 2023

Preprint

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Global microplastic (MP) pollution is now well recognized, with humans and animals consuming and inhaling MPs on a daily basis. Herein we described the effects of azide-free, 1 µm polystyrene MP beads co-delivered into lungs with a SARS-CoV-2 omicron BA.5 inoculum using a mouse model of mild COVID-19. Lung virus titres and viral RNA levels were not significantly affected by MPs, with overt clinical or histopathological changes also not observed. However, RNA-Seq of infected lungs revealed that MP exposure suppressed innate immune responses at 2 days post infection (dpi) and increased pro-inflammatory signatures at 6 dpi. The cytokine profile at 6 dpi showed a significant correlation with the ‘cytokine release syndrome’ signature seen in some severe COVID-19 patients. This study adds to a growing body of literature suggesting that MPs can dysregulate inflammation in specific disease settings.Graphical AbstractHIGHLIGHTSA single inoculation of microplastics dysregulated SARS-CoV-2 lung inflammationAt the peak of SARS-CoV-2 infection microplastics decreased early innate responsesLater post infection microplastics promoted a “cytokine release syndrome” signatureA key mechanism may involve the inhibition of the phagocytosis of infected cellsAzide-free microplastics were used, with no elevated ROS responses identifiedPostulated mechanisms whereby microplastics might decrease the proinflammatory responses 2 days after SARS-CoV-2 infection, yet promote the proinflammatory ‘cytokine release syndrome’ signature at 6 days post infection.

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“…hBOs were reprogrammed from adult dermal fibroblasts (HDFa, Gibco, C0135C) using the CytoTune-iPS 2.0 Sendai Reprogramming Kit (Invitrogen, A16518) (31), and were grown using the CelVivo Clinostar incubator (Invitro Technologies) as described (32). On the day of infection, ~30-day-old hBOs were transferred from each Clinoreactor into an ultra-low-binding 24 well plate (one hBO per well), and each hBO was infected with 10 5 CCID 50 of either JEV Nakayama , JEV FU , JEV NSW/22 , MVEV TC123130 , Imojev (33), or YFV 17D (30) for ~4 hrs.…”

Section: Infection Of Human Cortical Brain Organoidsmentioning

confidence: 99%

Neurovirulence of the Australian outbreak Japanese Encephalitis virus genotype 4 is lower compared to genotypes 2 and 3 in mice and human cortical brain organoids

Nguyen

Stewart

Tang

et al. 2023

Preprint

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Human infections with Japanese encephalitis virus (JEV) are a leading cause of viral encephalitis, with ≈70,000 symptomatic cases (≈40% with long-term neurological sequelae) and ≈20,000 deaths reported annually, primarily in Asia and the Western Pacific. An outbreak of JEV genotype 4 was also recently reported in Australia, with an isolate (JEVNSW/22) obtained from a stillborn piglet. Herein we characterize the neuropathology of JEVNSW/22, JEVFU (genotype 2) and JEVNakayama (genotype 3) in adult C57BL/6J wild-type mice, mice deficient in interferon regulatory factor 7 (IRF7-/-), and mice deficient in the type I interferon receptor (IFNAR-/-). In C57BL/6J and IRF7-/- mice with lethal outcomes, viral antigen was detected by immunohistochemistry in inter alia the cortex, thalamus and hippocampus. In these mice, histological lesions included neuronal degeneration, neuronal vacuolation, perivascular cuffing, leukocyte infiltrates, hemorrhage, and microgliosis, with apoptosis and astrocyte activation detected by immunohistochemistry. Microgliosis and hemorrhage persisted in some surviving mice ≈3-4 weeks post infection. JEV was universally lethal in IFNAR-/ mice by day 3 with histological signs of brain hemorrhage, but produced no other detectable brain infection or lesions, with NS1 readily detected in blood vessels, but not neurons, by immunohistochemistry. All JEV isolates showed robust productive cytopathic infection of human cortical brain organoids (hBOs), as well as a human neural progenitor cell line (RENcell VM). We thus describe a new IRF7-/-mouse model for JEV, which shows increased penetrance of lethal neuroinvasiveness and recapitulates many aspects human disease. Although less virulent in IRF7-/- mice, JEVNSW/22 retained the capacity to cause lethal encephalitis and to replicate and destroy human neuronal cells.

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Increased neurovirulence of omicron BA.5 and XBB variants over BA.1 in K18-hACE2 mice and human brain organoids

Cited by 5 publications

References 182 publications

Characterisation of a Japanese Encephalitis virus genotype 4 isolate from the 2022 Australian outbreak

Characterisation of a Japanese Encephalitis virus genotype 4 isolate from the 2022 Australian outbreak

Microplastics dysregulate innate immunity in the SARS-CoV-2 infected lung

Neurovirulence of the Australian outbreak Japanese Encephalitis virus genotype 4 is lower compared to genotypes 2 and 3 in mice and human cortical brain organoids

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