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1.
Sci Rep ; 11(1): 16201, 2021 08 10.
Article in English | MEDLINE | ID: covidwho-1351977

ABSTRACT

Optical spectroscopic techniques have been commonly used to detect the presence of biofilm-forming pathogens (bacteria and fungi) in the agro-food industry. Recently, near-infrared (NIR) spectroscopy revealed that it is also possible to detect the presence of viruses in animal and vegetal tissues. Here we report a platform based on visible and NIR (VNIR) hyperspectral imaging for non-contact, reagent free detection and quantification of laboratory-engineered viral particles in fluid samples (liquid droplets and dry residue) using both partial least square-discriminant analysis and artificial feed-forward neural networks. The detection was successfully achieved in preparations of phosphate buffered solution and artificial saliva, with an equivalent pixel volume of 4 nL and lowest concentration of 800 TU·[Formula: see text]L-1. This method constitutes an innovative approach that could be potentially used at point of care for rapid mass screening of viral infectious diseases and monitoring of the SARS-CoV-2 pandemic.


Subject(s)
Image Processing, Computer-Assisted/methods , Lentivirus Infections/diagnosis , Molecular Diagnostic Techniques/methods , Spectroscopy, Near-Infrared/methods , HEK293 Cells , Humans , Image Processing, Computer-Assisted/standards , Lentivirus/isolation & purification , Lentivirus/pathogenicity , Lentivirus Infections/virology , Molecular Diagnostic Techniques/standards , Point-of-Care Systems , Saliva/virology , Sensitivity and Specificity , Spectroscopy, Near-Infrared/standards
2.
FASEB J ; 35(9): e21801, 2021 09.
Article in English | MEDLINE | ID: covidwho-1345745

ABSTRACT

The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays a crucial role in mediating viral entry into host cells. However, whether it contributes to pulmonary hyperinflammation in patients with coronavirus disease 2019 is not well known. In this study, we developed a spike protein-pseudotyped (Spp) lentivirus with the proper tropism of the SARS-CoV-2 spike protein on the surface and determined the distribution of the Spp lentivirus in wild-type C57BL/6J male mice that received an intravenous injection of the virus. Lentiviruses with vesicular stomatitis virus glycoprotein (VSV-G) or with a deletion of the receptor-binding domain (RBD) in the spike protein [Spp (∆RBD)] were used as controls. Two hours postinfection (hpi), there were 27-75 times more viral burden from Spp lentivirus in the lungs than in other organs; there were also about 3-5 times more viral burden from Spp lentivirus than from VSV-G lentivirus in the lungs, liver, kidney, and spleen. Deletion of RBD diminished viral loads in the lungs but not in the heart. Acute pneumonia was observed in animals 24 hpi. Spp lentivirus was mainly found in SPC+ and LDLR+ pneumocytes and macrophages in the lungs. IL6, IL10, CD80, and PPAR-γ were quickly upregulated in response to infection in the lungs as well as in macrophage-like RAW264.7 cells. Furthermore, forced expression of the spike protein in RAW264.7 cells significantly increased the mRNA levels of the same panel of inflammatory factors. Our results demonstrated that the spike protein of SARS-CoV-2 confers the main point of viral entry into the lungs and can induce cellular pathology. Our data also indicate that an alternative ACE2-independent viral entry pathway may be recruited in the heart and aorta.


Subject(s)
Macrophages/immunology , Pneumonia, Viral/immunology , Pneumonia, Viral/pathology , Spike Glycoprotein, Coronavirus/immunology , Acute Disease , Alveolar Epithelial Cells/virology , Animals , B7-1 Antigen , Cell Line , Inflammation Mediators , Interleukin-10 , Interleukin-6 , Lentivirus/genetics , Lentivirus/isolation & purification , Lentivirus/metabolism , Lung/immunology , Lung/pathology , Lung/virology , Macrophages/virology , Male , Membrane Glycoproteins , Mice , Mice, Inbred C57BL , PPAR gamma , RAW 264.7 Cells , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/genetics , Spike Glycoprotein, Coronavirus/metabolism , Viral Envelope Proteins
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