Pseudotyped GFP rSARS-CoV-2 Spike D614G (COVG-D641G)

Product Overview
SARS-CoV-2 (D614G) Pseudovirus are used to test the ability of serum, antibodies, and drugs to neutralize the infectivity of SARS-CoV-2 spike protein. Pseudovirus display antigenically correct spike protein pseudotyped on replication-incompetent virus particles that contain a heterologous lentiviral (HIV) core. Pseudovirus are capable of a single round of infection and carry a genome that expresses GFP reporter gene upon infection. Pseudovirus are produced in HEK293T cells using three separate plasmids, encoding the spike protein, a lentiviral gag polyprotein, and a reporter gene. Pseudovirus are created using a second-generation lentiviral system with components that are highly unlikely to recombine to produce a fully infectious virus (requiring 3 separate recombination events to do so). However, lentiviruses are capable of genomic integration and Pseudovirus are derived from biological materials so should be handled with caution within a BSL2 or enhanced BSL2 laboratory environment.
Nature
Virus
Application Notes
We recommended to use 10-20 uL pseudotyped virus per 1E+04 293T cells for in vitro assay. Due to differences in cell status,the best infection conditions and MOI should be determined by the end user.The virus can be diluted with cell culture medium if needed.
Size
1 mL
Storage
Store at -80°C. Multiple freeze/thaw cycles not recommended.
When using the virus, transfer the virus from the -80 ° C refrigerator and melt it in an ice bath.
Ship
Frozen on dry ice
Warnings
Biosafety Level:    BSL-2
It is the responsibility of the principal investigator to seek Institutional Biosafety Safety Committee approval for recombinant DNA, transgenic animal or infectious agent use within their laboratory spaces and maintain an Institutional Biosafety Safety Committee approval during the time period these materials are used.

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References


Corona in Children: the Co-Ki Study Relevance of SARS-CoV-2 in outpatient pediatric services in Germany

MONATSSCHRIFT KINDERHEILKUNDE

Authors: Schwarz, Silke; Jenetzky, Ekkehart; Krafft, Hanno; Maurer, Tobias; Steuber, Christian; Reckert, Till; Fischbach, Thomas; Martin, David

Background. In Germany over 80% of children and adolescents are in the ambulatory care of registered pediatricians. These have a specific perspective on the COVID-19 pandemic. Methods. For this reason, this professional group initiated a central recording of case numbers, individual case descriptions and observations on infections and illnesses with SARS-CoV-2 (www.co-ki.de). Results. So far 557 pediatricians have participated. Together they care for ca. 670,000 children. They reported 9803 children who presented as suspected cases. The pediatricians themselves had a clinical suspicion of SARS-CoV-2 infections in 3654 children. In 7707 children PCR tests were carried out using nose/throat swabs of which 198 (2.6%) were positive. In addition, 731 children were tested for SARS-CoV-2 antibodies with detection in 82 cases (11.2%). Despite initially positive PCR tests, 47 children had a negative antibody test at least 2 weeks later. Our query as to infections of adults by children yielded only one case, which a telephone enquiry revealed as unlikely. Discussion. From an outpatient pediatric perspective COVID-19 is rare. There was no convincing evidence that children are a relevant source of infection for SARS-CoV-2 nor that they are relevantly at risk.

Altered bioenergetics and mitochondrial dysfunction of monocytes in patients with COVID-19 pneumonia

EMBO MOLECULAR MEDICINE

Authors: Gibellini, Lara; De Biasi, Sara; Paolini, Annamaria; Borella, Rebecca; Boraldi, Federica; Mattioli, Marco; Lo Tartaro, Domenico; Fidanza, Lucia; Caro-Maldonado, Alfredo; Meschiari, Marianna; Iadisernia, Vittorio; Bacca, Erica; Riva, Giovanni; Cicchetti, Luca; Quaglino, Daniela; Guaraldi, Giovanni; Busani, Stefano; Girardis, Massimo; Mussini, Cristina; Cossarizza, Andrea

In patients infected by SARS-CoV-2 who experience an exaggerated inflammation leading to pneumonia, monocytes likely play a major role but have received poor attention. Thus, we analyzed peripheral blood monocytes from patients with COVID-19 pneumonia and found that these cells show signs of altered bioenergetics and mitochondrial dysfunction, had a reduced basal and maximal respiration, reduced spare respiratory capacity, and decreased proton leak. Basal extracellular acidification rate was also diminished, suggesting reduced capability to perform aerobic glycolysis. Although COVID-19 monocytes had a reduced ability to perform oxidative burst, they were still capable of producing TNF and IFN-gamma in vitro. A significantly high amount of monocytes had depolarized mitochondria and abnormal mitochondrial ultrastructure. A redistribution of monocyte subsets, with a significant expansion of intermediate/pro-inflammatory cells, and high amounts of immature monocytes were found, along with a concomitant compression of classical monocytes, and an increased expression of inhibitory checkpoints like PD-1/PD-L1. High plasma levels of several inflammatory cytokines and chemokines, including GM-CSF, IL-18, CCL2, CXCL10, and osteopontin, finally confirm the importance of monocytes in COVID-19 immunopathogenesis.

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