The pseudovirus technology platform of NIFDC plays an important role in the prevention and control of SARS-CoV-2.
1. NIFDC established a robust pseudovirus-based neutralization assay for SARS-CoV-2 and shared pseudoviruses and related protocols with the developers of vaccines or therapeutics to fight against this lethal virus form February 2020 to now. https://doi.org/10.1080/22221751.2020.1743767
2. The SOP for “Quantification of SARS-CoV-2 neutralizing antibody by a pseudotyped virus-based assay”. https://doi.org/10.1038/s41596-020-0394-5
3. They found that the dominant D614G itself and combined with other mutations are more infectious, which was published in Cell(2020). In this study, Over 100 mutations were selected for analyses on their infectivity and antigenicity. Ablation of both N331 and N343 glycosylation at RBD drastically reduced infectivity. Ten mutations such as N234Q, L452R, A475V, and V483A was markedly resistant to some mAbs. https://doi.org/10.1016/j.cell.2020.07.012
4. The origin and intermediate host for SARS-CoV-2 is yet to be determined. Coronaviruses found to be closely related to SARS-CoV-2 include RaTG13 derived from bat and two clusters (PCoV-GD and PCoV-GX) of coronaviruses identified in pangolin. Here, we studied the infectivity and antigenicity patterns of SARS-CoV-2 and the three related coronaviruses. Compared with the other three viruses, RaTG13 showed almost no infectivity to a variety of cell lines. The two pangolin coronaviruses and SARS-CoV-2 showed similar infectious activity. However, in SARS-CoV-2-susceptible cell lines, the pangolin coronaviruses presented even higher infectivity. The striking difference between the SARS-CoV-2 and pangolin coronaviruses is that the latter can infect porcine cells, which could be partially attributed to an amino acid difference at the position of 498 of the spike protein. The infection by SARS-CoV-2 was mainly mediated by Furin and TMPRSS2, while PCoV-GD and PCoV-GX mainly depend on Cathepsin L. Extensive cross-neutralization was found between SARS-CoV-2 and PCoV-GD. However, almost no cross-neutralization was observed between PCoV-GX and SARS-CoV-2 or PCoV-GD. More attention should be paid to pangolin coronaviruses and to investigate the possibility of these coronaviruses spreading across species to become zoonoses among pigs or humans. https://doi.org/10.1038/s41421-021-00256-3
5. More than 300 strains of sars-cov-2 with different mutations were constructed by using pseudovirus technology. The mutant pseudovirus library was used to evaluate the infection and antigenicity of mutant viral strains:
We found no evidence that mink variants have altered tropisms compared to the currently predominant D614G variant. Moreover, compared to the currently predominant D614G variant. Moreover, 69-70 del variants did not appear to become resistant to RBD-specific mAb and polyclonal antibodies from animals and convalescent patients. It is of note that the recently surging variants of B.1.1.7 lineage in the UK carry multiple mutations including the 69-70 del studied here. More studies would be needed to better understand the impact of 69-70 del mutation on the transmission and antigenicity of the B.1.1.7 variants. Furthermore, the decreased sensitivity of the F486L variants to some neutralizing mAb and selected convalescent plasma warrants further investigations, given the implication on the formulation of mAb-based antiviral therapies. https://doi.org/10.1038/s41392-021-00617-0.
The 501Y.V2 variants of SARS-CoV-2 containing multiple mutations in spike are now dominant in South Africa and are rapidly spreading to other countries. Here, experiments with 18 pseudotyped viruses showed that the 501Y.V2 variants do not confer increased infectivity in multiple cell types except for murine ACE2-overexpressing cells, where a substantial increase in infectivity was observed. Notably, the susceptibility of the 501Y.V2 variants to 12 of 17 neutralizing monoclonal antibodies was substantially diminished, and the neutralization ability of the sera from convalescent patients and immunized mice was also reduced for these variants. The neutralization resistance was mainly caused by E484K and N501Y mutations in the receptorbinding domain of spike. The enhanced infectivity in murine ACE2-overexpressing cells suggests the possibility of spillover of the 501Y.V2 variants to mice. Moreover, the neutralization resistance we detected for the 501Y.V2 variants suggests the potential for compromised efficacy of monoclonal antibodies and vaccines. https://doi.org/10.1016/j.cell.2021.02.042
6. The detection technology of pseudovirus based neutralization antibody has been widely used in vaccine evaluation( mRNA vaccine: https://doi.org/10.1016/j.cell.2020.07.024; RBD-subunit vaccines: https://doi.org/10.1038/s41422-021-00514-9, https://doi.org/10.1038/s41467-021-21634-1 etc.), Therapeutic monoclonal antibody research and development evaluation(https://doi.org/10.1038/s41467-020-19568-1, https://science.sciencemag.org/content/369/6510/1505, etc), drug research and development( https://doi.org/10.1038/s41392-021-00558-8 ), seroepidemiological investigation of infected people(https://doi.org/10.1038/s41564-020-00824-5, https://doi.org/10.1007/s11684-020-0822-5 ), etc.
link:https://www.nifdc.org.cn//nifdc/gzdt/ywdt/20200821084200953.html
