EBioMedicine. 2021 Dec 31
Jianxiong Zeng, Xiaochun Xie, Xiao-Li Feng, Ling Xu,Jian-Bao Han, Dandan Yu, Qing-Cui Zou, Qianjin Liu, Xiaohong Li, Guanqin Ma, Ming-Hua Li, and Yong-Gang Yao
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AAV vector packaging For AAV-hACE2 infection in mice, C57BL/6J (n = 5) and Nlrp3-KO (n = 5) mice were intranasally incubated with hACE2-expressing AAV (AAV-hACE2) (PackGene Biotech, Guangzhou) at a total of 40 ml containing 4 x10^12 viral genome copies (GC) (20 ml per nostril). Request Quote

Research Field: Covid-related

Keywords: SARS-CoV-2, NLRP3 inflammasome, Immune overactivation, Cytokine storm; MCC950

Dose: 4 x10^12 viral genome copies

Routes of Administration: intranasally incubated

Animal or cell line strain: C57BL/6J (n = 5) and Nlrp3-KO (n = 5) mice


Background: The Coronavirus Disease 2019 (COVID-19) pandemic has been a great threat to global public health since 2020. Although the advance on vaccine development has been largely achieved, a strategy to alleviate immune overactivation in severe COVID-19 patients is still needed. The NLRP3 inflammasome is activated upon SARS-CoV-2 infection and associated with COVID-19 severity. However, the processes by which the NLRP3 inflammasome is involved in COVID-19 disease remain unclear.
Methods: We infected THP-1 derived macrophages, NLRP3 knockout mice, and human ACE2 transgenic mice with live SARS-CoV-2 in Biosafety Level 3 (BSL-3) laboratory. We performed quantitative real-time PCR for targeted viral or host genes from SARS-CoV-2 infected mouse tissues, conducted histological or immunofluorescence analysis in SARS-CoV-2 infected mouse tissues. We also injected intranasally AAV-hACE2 or intraperitoneally NLRP3 inflammasome inhibitor MCC950 before SARS-CoV-2 infection in mice as indicated.
Findings: We have provided multiple lines of evidence that the NLRP3 inflammasome plays an important role in the host immune response to SARS-CoV-2 invasion of the lungs. Inhibition of the NLRP3 inflammasome attenuated the release of COVID-19 related pro-inflammatory cytokines in cell cultures and mice. The severe pathology induced by SARS-CoV-2 in lung tissues was reduced in Nlrp3−/− mice compared to wild-type C57BL/6 mice. Finally, specific inhibition of the NLRP3 inflammasome by MCC950 alleviated excessive lung inflammation and thus COVID-19 like pathology in human ACE2 transgenic mice.

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