Highly pathogenic coronavirus N protein aggravates lung injury by MASP-2-mediated complement over-activation

Ting Gao; Mingdong Hu; Xiaopeng Zhang; Hongzhen Li; Lin Zhu; Hainan Liu; Qincai Dong; Zhang Zhang; Zhongyi Wang; Yong Hu; Yangbo Fu; Yanwen Jin; Kaitong Li; Songtao Zhao; Yongjiu Xiao; Shuping Luo; Lufeng Li; Lingfang Zhao; Junli Liu; Huailong Zhao; Yue Liu; Weihong Yang; Jing Peng; Xiaoyu Chen; Ping Li; Yaoning Liu; Yonghong Xie; Jibo Song; Lu Zhang; Qingjun Ma; Xiuwu Bian; Wei Chen; Xuan Liu; Qing Mao; Cheng Cao

The copyright holder for this preprint (which was not peer-reviewed) is . https://doi.org/10.1101/2020.03. 29.20041962 doi: medRxiv preprint Detection of C5a in COVID-19 patients Sera from mild or severe COVID-19 patients were collected under the approval of hospital ethic committee. Server patients were defined as fever or suspected respiratory infection, plus one of respiratory rate >30 breaths/min, severe respiratory distress, or SpO2 <90% on room air. Patients with pneumonia and no signs of severe pneumonia are defined as mild cases. Sera 5 from 12 mild patients (6 males and 6 females), with an average of 5612.1 years old and an average 26.39.1 days of illness, and sera from 18 severe patients (7 males and 11 females), with an average of 72.410.1 years old and an average 40.1512.74 days of illness, were assayed. Sera from healthy people for physical were collected from the clinical laboratory. Serum C5a level was detected by double antibody sandwich ELISA (R&D Systems). 10 Anti-C5a antibody therapy BDB-001 is a mouse-human chimeric (IgG4) antibody against human C5a. It was manufactured from IFX-1 cell line by Staidson in accordance with the Good Manufacture Practices. The IFX-1 cell line has originally been developed by InflaRx GmbH, Germany and was licensed to Beijing Deferengrei Biotech Co., Ltd., a wholly owned subsidiary of Staidson 15 Biopharmaceutical Co., Ltd. The injection was approved by National Medical Product Administration (NMPA) of China for phase I clinical trial in healthy subjects in 2018. After the SARS-CoV-2 coronavirus induced pneumonia (COVID-19) outbreak, the antibody was approved by NMPA for phase II clinical trials for the treatment of COVID-19 on February 7, 2020 (2020L00003). Administration of the antibody to human was also approved by the Ethics 20 Committee in Wuhan Huoshenshan Hospital. 300 mg anti-C5a antibody in 250 ml saline was administrated i.v. on day 1, 2, 3, 5, 7, 9, 11 and 13. Arterial blood gas (ABG) test, C reaction protein (CRP), blood routine (blood RT) and Hepatic function were regularly determined. SaO2, blood pressure, heart beat were monitored as required. 25 The data were presented as the mean ±SD or the mean ±SEM and analyzed using the two-tailed Student t test or ANOVA. Survival curves were analyzed by Log-rank test, Gehan-Breslow-Wilcoxon test or parametric regression model. Differences were considered significant at *P <0.05, **P <0.01, or ***P<0.001 as indicated. the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.