| Autor: |
Bullen CK; Johns Hopkins University, School of Medicine, Department of Medicine, Division of Infectious Diseases, Baltimore, MD, USA., Hogberg HT; Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, MD, USA., Bahadirli-Talbott A; Johns Hopkins University, School of Medicine, Department of Medicine, Division of Infectious Diseases, Baltimore, MD, USA., Bishai WR; Johns Hopkins University, School of Medicine, Department of Medicine, Division of Infectious Diseases, Baltimore, MD, USA., Hartung T; Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, MD, USA.; CAAT-Europe, University of Konstanz, Konstanz, Germany.; Johns Hopkins University, Bloomberg School of Public Health, Department of Molecular Microbiology and Immunology, Baltimore, MD, USA., Keuthan C; Johns Hopkins University, School of Medicine, Department of Ophthalmology, Wilmer Eye Institute, Baltimore, MD, USA., Looney MM; Johns Hopkins University, School of Medicine, Department of Medicine, Division of Infectious Diseases, Baltimore, MD, USA., Pekosz A; Johns Hopkins University, Bloomberg School of Public Health, Department of Molecular Microbiology and Immunology, Baltimore, MD, USA., Romero JC; Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, MD, USA., Sillé FCM; Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, MD, USA.; Johns Hopkins University, Bloomberg School of Public Health, Department of Environmental Health & Engineering, Baltimore, MD, USA., Um P; Johns Hopkins University, School of Medicine, Department of Medicine, Division of Infectious Diseases, Baltimore, MD, USA., Smirnova L; Johns Hopkins University, Bloomberg School of Public Health, Center for Alternatives to Animal Testing (CAAT), Baltimore, MD, USA. |
| Abstrakt: |
Reports from Wuhan suggest that 36% of COVID-19 patients show neurological symptoms, and cases of viral encephalitis have been reported, suggesting that the virus is neurotropic under unknown circumstances. This is well established for other coronaviruses. In order to understand why some patients develop such symptoms and others do not, we address herein the infectability of the central nervous system (CNS). Reports that the ACE2 receptor – critical for virus entry into lung cells – is found in different neurons support this expectation. We employed a human induced pluripotent stem cell (iPSC)- derived BrainSphere model, which we used earlier for Zika, Dengue, HIV and John Cunningham virus infection studies. We detected the expression of the ACE2 receptor, but not TMPRSS2, in the model. Incubating the BrainSpheres for 6 hours with SARS-CoV-2 at a multiplicity of infection (MOI) of 0.1 led to infection of a fraction of neural cells with replication of the virus evident at 72 hpi. Virus particles were found in the neuronal cell body extending into apparent neurite structures. PCR measurements corroborated the replication of the virus, suggesting at least a tenfold increase in virus copies per total RNA. Leveraging state-of-the-art 3D organotypic cell culture, which has been shown to allow both virus infection and modeling of (developmental) neurotoxicity but is at the same time simple enough to be transferred and used in a BSL-3 environment, we demonstrate, for the first time, the potential critically important neurotropism of SARS-CoV-2. |
