tetano
Editor, Senior Moderator
Front Immunol
. 2021 Dec 7;12:791753.
doi: 10.3389/fimmu.2021.791753. eCollection 2021.
SARS-CoV-2 N Protein Induces Acute Lung Injury in Mice via NF-ĸB Activation
Jie Xia[SUP] 1 [/SUP], Wenqi Tang[SUP] 1 [/SUP], Jiangmei Wang[SUP] 1 [/SUP], Dengming Lai[SUP] 1 [/SUP], Qi Xu[SUP] 2 [/SUP], Ruoqiong Huang[SUP] 1 [/SUP], Yaoqin Hu[SUP] 1 [/SUP], Xiaojue Gong[SUP] 1 [/SUP], Jiajie Fan[SUP] 1 [/SUP], Qiang Shu[SUP] 1 [/SUP], Jianguo Xu[SUP] 1 [/SUP]
Affiliations
Abstract
Background: Infection of SARS-CoV-2 may cause acute respiratory syndrome. It has been reported that SARS-CoV-2 nucleocapsid protein (N-protein) presents early in body fluids during infection. The direct involvement of N-protein in lung injury is poorly understood.
Methods: Recombinant N-protein was pretreated with polymyxin B, a lipopolysaccharide (LPS)-neutralizing agent. C57BL/6, C3H/HeJ (resistant to LPS), and C3H/HeN (control for C3H/HeJ) mice were exposed to N-protein via intratracheal administration to examine acute lung injury. In vitro, bone marrow-derived macrophages (BMDMs) were cultured with N-protein to study phosphorylation of nuclear factor kappa B (NF-ĸB) p65, macrophage polarization, and expression of proinflammatory cytokines.
Results: N-protein produced acute lung injury in C57BL/6 mice, with elevated protein permeability, total cell count, neutrophil infiltration, and proinflammatory cytokines in the bronchioalveolar lavage. N-protein also induced lung injury in both C3H/HeJ and C3H/HeN mice, indicating that the effect could not be attributed to the LPS contamination. N-protein triggered phosphorylation of NF-ĸB p65 in vitro, which was abolished by both N-protein denaturation and treatment with an antibody for N-protein, demonstrating that the effect is N-protein specific. In addition, N-protein promoted M1 macrophage polarization and the expression of proinflammatory cytokines, which was also blocked by N-protein denaturation and antibody for N-protein. Furthermore, N-protein induced NF-ĸB p65 phosphorylation in the lung, while pyrrolidine dithiocarbamate, an NF-ĸB inhibitor, alleviated the effect of N-protein on acute lung injury.
Conclusions: SARS-CoV-2 N-protein itself is toxic and induces acute lung injury in mice. Both N-protein and NF-ĸB pathway may be therapeutic targets for treating multi-organ injuries in Coronavirus disease 2019 (COVID-19).
Keywords: COVID-19; NF- kappa B; SARS-CoV-2; acute lung injury; nucleocapsid (N) protein.
. 2021 Dec 7;12:791753.
doi: 10.3389/fimmu.2021.791753. eCollection 2021.
SARS-CoV-2 N Protein Induces Acute Lung Injury in Mice via NF-ĸB Activation
Jie Xia[SUP] 1 [/SUP], Wenqi Tang[SUP] 1 [/SUP], Jiangmei Wang[SUP] 1 [/SUP], Dengming Lai[SUP] 1 [/SUP], Qi Xu[SUP] 2 [/SUP], Ruoqiong Huang[SUP] 1 [/SUP], Yaoqin Hu[SUP] 1 [/SUP], Xiaojue Gong[SUP] 1 [/SUP], Jiajie Fan[SUP] 1 [/SUP], Qiang Shu[SUP] 1 [/SUP], Jianguo Xu[SUP] 1 [/SUP]
Affiliations
- PMID: 34950152
- PMCID: PMC8688532
- DOI: 10.3389/fimmu.2021.791753
Abstract
Background: Infection of SARS-CoV-2 may cause acute respiratory syndrome. It has been reported that SARS-CoV-2 nucleocapsid protein (N-protein) presents early in body fluids during infection. The direct involvement of N-protein in lung injury is poorly understood.
Methods: Recombinant N-protein was pretreated with polymyxin B, a lipopolysaccharide (LPS)-neutralizing agent. C57BL/6, C3H/HeJ (resistant to LPS), and C3H/HeN (control for C3H/HeJ) mice were exposed to N-protein via intratracheal administration to examine acute lung injury. In vitro, bone marrow-derived macrophages (BMDMs) were cultured with N-protein to study phosphorylation of nuclear factor kappa B (NF-ĸB) p65, macrophage polarization, and expression of proinflammatory cytokines.
Results: N-protein produced acute lung injury in C57BL/6 mice, with elevated protein permeability, total cell count, neutrophil infiltration, and proinflammatory cytokines in the bronchioalveolar lavage. N-protein also induced lung injury in both C3H/HeJ and C3H/HeN mice, indicating that the effect could not be attributed to the LPS contamination. N-protein triggered phosphorylation of NF-ĸB p65 in vitro, which was abolished by both N-protein denaturation and treatment with an antibody for N-protein, demonstrating that the effect is N-protein specific. In addition, N-protein promoted M1 macrophage polarization and the expression of proinflammatory cytokines, which was also blocked by N-protein denaturation and antibody for N-protein. Furthermore, N-protein induced NF-ĸB p65 phosphorylation in the lung, while pyrrolidine dithiocarbamate, an NF-ĸB inhibitor, alleviated the effect of N-protein on acute lung injury.
Conclusions: SARS-CoV-2 N-protein itself is toxic and induces acute lung injury in mice. Both N-protein and NF-ĸB pathway may be therapeutic targets for treating multi-organ injuries in Coronavirus disease 2019 (COVID-19).
Keywords: COVID-19; NF- kappa B; SARS-CoV-2; acute lung injury; nucleocapsid (N) protein.