Quantitative proteomic analysis of SARS-CoV-2 infection of primary human airway ciliated cells and lung epithelial cells demonstrates the effectiveness of SARS-CoV-2 innate immune evasion

Wellcome Open Research

article

Thomas W.M. Crozier¹  Edward J.D. Greenwood¹  James C. Williamson¹  Wenrui Guo²  Linsey M. Porter²  Ildar Gabaev¹  Ana Teixeira-Silva¹  Guinevere L. Grice¹  Arthur Wickenhagen³  Richard J. Stanton⁴  Eddie C. Y. Wang⁴  Sam J. Wilson³  Nicholas J. Matheson¹  James A. Nathan¹  Frank McCaughan²  Paul J. Lehner¹ 

[1] Department of Medicine, Cambridge Institute of Therapeutic Immunology and Infectious Disease, University of Cambridge;Department of Medicine, Addenbrookes Hospital, University of Cambridge;MRC - University of Glasgow Centre for Virus Research;Division of Infection and Immunity, School of Medicine, Cardiff University;NHS Blood and Transplant

关键词: SARS-CoV-2;    COVID-19;    Coronavirus;    proteomics;   

DOI  :  10.12688/wellcomeopenres.17946.1

学科分类：内科医学

来源: Wellcome

【 摘 要 】

Background: Quantitative proteomics is able to provide a comprehensive, unbiased description of changes to cells caused by viral infection, but interpretation may be complicated by differential changes in infected and uninfected ‘bystander’ cells, or the use of non-physiological cellular models.Methods: In this paper, we use fluorescence-activated cell sorting (FACS) and quantitative proteomics to analyse cell-autonomous changes caused by authentic SARS-CoV-2 infection of respiratory epithelial cells, the main target of viral infectionin vivo. First, we determine the relative abundance of proteins in primary human airway epithelial cells differentiated at the air-liquid interface (basal, secretory and ciliated cells). Next, we specifically characterise changes caused by SARS-CoV-2 infection of ciliated cells. Finally, we compare temporal proteomic changes in infected and uninfected ‘bystander’ Calu-3 lung epithelial cells and compare infection with B.29 and B.1.1.7 (Alpha) variants.Results: Amongst 5,709 quantified proteins in primary human airway ciliated cells, the abundance of 226 changed significantly in the presence of SARS-CoV-2 infection (q 1.5-fold). Notably, viral replication proceeded without inducing a type-I interferon response. Amongst 6,996 quantified proteins in Calu-3 cells, the abundance of 645 proteins changed significantly in the presence of SARS-CoV-2 infection (q 1.5-fold). In contrast to the primary cell model, a clear type I interferon (IFN) response was observed. Nonetheless, induction of IFN-inducible proteins was markedly attenuated in infected cells, compared with uninfected ‘bystander’ cells. Infection with B.29 and B.1.1.7 (Alpha) variants gave similar results.Conclusions: Taken together, our data provide a detailed proteomic map of changes in SARS-CoV-2-infected respiratory epithelial cells in two widely used, physiologically relevant models of infection. As well as identifying dysregulated cellular proteins and processes, the effectiveness of strategies employed by SARS-CoV-2 to avoid the type I IFN response is illustrated in both models.

【 授权许可】

CC BY