SARS-CoV replicates in primary human alveolar type II cell cultures but not in type I-like cells

Abstract Severe acute respiratory syndrome (SARS) is a disease characterized by diffuse alveolar damage. We isolated human alveolar type II cells and maintained them in a highly differentiated state. Type II cell cultures supported SARS-CoV replication as evidenced by RT-PCR detection of viral subgenomic RNA and an increase in virus titer. Virus titers were maximal by 24 h and peaked at approximately 105 pfu/mL. Two cell types within the cultures were infected. One cell type was type II cells, which were positive for SP-A, SP-C, cytokeratin, a type II cell-specific monoclonal antibody, and Ep-CAM. The other cell type was composed of spindle-shaped cells that were positive for vimentin and collagen III and likely fibroblasts. Viral replication was not detected in type I-like cells or macrophages. Hence, differentiated adult human alveolar type II cells were infectible but alveolar type I-like cells and alveolar macrophages did not support productive infection.

[1]  S. Asa,et al.  Pulmonary pathology of severe acute respiratory syndrome in Toronto , 2005, Modern Pathology.

[2]  M. McNutt,et al.  Molecular Pathology in the Lungs of Severe Acute Respiratory Syndrome Patients , 2007, The American Journal of Pathology.

[3]  R. Mason Biology of alveolar type II cells , 2006, Respirology.

[4]  C. Schwegmann-Wessels,et al.  Analysis of ACE2 in polarized epithelial cells: surface expression and function as receptor for severe acute respiratory syndrome-associated coronavirus. , 2006, The Journal of general virology.

[5]  N. Pedemonte,et al.  Contribution of CFTR to apical-basolateral fluid transport in cultured human alveolar epithelial type II cells. , 2006, American journal of physiology. Lung cellular and molecular physiology.

[6]  K. Subbarao,et al.  Is there an ideal animal model for SARS? , 2006, Trends in Microbiology.

[7]  J. Crapo,et al.  Allometric relationships of cell numbers and size in the mammalian lung. , 1992, American journal of respiratory cell and molecular biology.

[8]  S. Perlman,et al.  ACE2 Receptor Expression and Severe Acute Respiratory Syndrome Coronavirus Infection Depend on Differentiation of Human Airway Epithelia , 2005, Journal of Virology.

[9]  Junya Fukuoka,et al.  Lung pathology of severe acute respiratory syndrome (SARS): a study of 8 autopsy cases from Singapore , 2003, Human Pathology.

[10]  Jill Taylor,et al.  Discovery of Novel Human and Animal Cells Infected by the Severe Acute Respiratory Syndrome Coronavirus by Replication-Specific Multiplex Reverse Transcription-PCR , 2004, Journal of Clinical Microbiology.

[11]  J. Druce,et al.  SARS–associated Coronavirus Replication in Cell Lines , 2006, Emerging infectious diseases.

[12]  J. Peiris,et al.  Academic Editor: Sherif Zaki, Centers for Disease Control, United States of America , 2005 .

[13]  N. Pedemonte,et al.  Erratum: Contribution of CFTR to apical-basolateral fluid transport in cultured human alveolar epithelial type II cells (American Journal of Physiology - Lung Cellular and Molecular Physiology (February 2006) 290, 34, (L242-L249)) , 2006 .

[14]  K. Brown,et al.  TNF-alpha sensitizes normal and fibrotic human lung fibroblasts to Fas-induced apoptosis. , 2006, American journal of respiratory cell and molecular biology.

[15]  A. Danchin,et al.  The Severe Acute Respiratory Syndrome , 2003 .

[16]  R. Wiener,et al.  Angiotensin converting enzyme 2 is primarily epithelial and is developmentally regulated in the mouse lung , 2007, Journal of cellular biochemistry.

[17]  R. Blumenthal,et al.  Human β-Defensins Suppress Human Immunodeficiency Virus Infection: Potential Role in Mucosal Protection , 2005, Journal of Virology.

[18]  Barry H. Smith,et al.  A continuous tumor‐cell line from a human lung carcinoma with properties of type II alveolar epithelial cells , 1976, International journal of cancer.

[19]  Y. Guan,et al.  Cytokine Responses in Severe Acute Respiratory Syndrome Coronavirus-Infected Macrophages In Vitro: Possible Relevance to Pathogenesis , 2005, Journal of Virology.

[20]  Krishna Shankara Narayanan,et al.  Exogenous ACE2 Expression Allows Refractory Cell Lines To Support Severe Acute Respiratory Syndrome Coronavirus Replication , 2005, Journal of Virology.

[21]  C. Peters,et al.  Interferon-beta and interferon-gamma synergistically inhibit the replication of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) , 2004, Virology.

[22]  John L. Sullivan,et al.  Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus , 2003, Nature.

[23]  Z. Li,et al.  Expression of elevated levels of pro‐inflammatory cytokines in SARS‐CoV‐infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS† , 2006, The Journal of pathology.

[24]  Xin Li,et al.  The clinical pathology of severe acute respiratory syndrome (SARS): a report from China , 2003, The Journal of pathology.

[25]  J. Crapo,et al.  Distribution of lung cell numbers and volumes between alveolar and nonalveolar tissue. , 1992, The American review of respiratory disease.

[26]  Q. Xie,et al.  RETRACTED: TGF-β1 induces alveolar epithelial to mesenchymal transition in vitro , 2004 .

[27]  Q. Xie,et al.  TGF-beta1 induces alveolar epithelial to mesenchymal transition in vitro. , 2004, Life sciences.

[28]  T. Ganz Antimicrobial polypeptides in host defense of the respiratory tract. , 2002, The Journal of clinical investigation.

[29]  Michelle M. Packard,et al.  Immunohistochemical, in situ hybridization, and ultrastructural localization of SARS-associated coronavirus in lung of a fatal case of severe acute respiratory syndrome in Taiwan , 2005, Human Pathology.

[30]  A. Nicholson,et al.  Induction of epithelial-mesenchymal transition in alveolar epithelial cells by transforming growth factor-beta1: potential role in idiopathic pulmonary fibrosis. , 2005, The American journal of pathology.

[31]  Yongsheng Chang,et al.  Differentiated human alveolar epithelial cells and reversibility of their phenotype in vitro. , 2007, American journal of respiratory cell and molecular biology.

[32]  Z. Lang,et al.  A clinicopathological study of three cases of severe acute respiratory syndrome (SARS) , 2003, Pathology.

[33]  K. Brown,et al.  TNF-α Sensitizes Normal and Fibrotic Human Lung Fibroblasts to Fas-Induced Apoptosis , 2006 .

[34]  G. Navis,et al.  Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis , 2004, The Journal of pathology.

[35]  Ralph S. Baric,et al.  Severe Acute Respiratory Syndrome Coronavirus Infection of Human Ciliated Airway Epithelia: Role of Ciliated Cells in Viral Spread in the Conducting Airways of the Lungs , 2005, Journal of Virology.

[36]  S. Harrison,et al.  SARS Coronavirus, but Not Human Coronavirus NL63, Utilizes Cathepsin L to Infect ACE2-expressing Cells , 2006, Journal of Biological Chemistry.

[37]  Jinhua Lu,et al.  The SARS coronavirus spike glycoprotein is selectively recognized by lung surfactant protein D and activates macrophages , 2007, Immunobiology.

[38]  Qingling Zhang,et al.  Organ distribution of severe acute respiratory syndrome (SARS) associated coronavirus (SARS‐CoV) in SARS patients: implications for pathogenesis and virus transmission pathways , 2004, The Journal of pathology.

[39]  Tsai-Jung Wu,et al.  Identification of pulmonary Oct-4+ stem/progenitor cells and demonstration of their susceptibility to SARS coronavirus (SARS-CoV) infection in vitro. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Yi Guan,et al.  Lung pathology of fatal severe acute respiratory syndrome , 2003, The Lancet.

[41]  Bo Zhang,et al.  Multiple organ infection and the pathogenesis of SARS , 2005, The Journal of experimental medicine.

[42]  M. Hynes Clinical Pathology , 1923 .

[43]  Lucy A. Perrone,et al.  Apical Entry and Release of Severe Acute Respiratory Syndrome-Associated Coronavirus in Polarized Calu-3 Lung Epithelial Cells , 2005, Journal of Virology.

[44]  G. Tse,et al.  Tissue and cellular tropism of the coronavirus associated with severe acute respiratory syndrome: an in‐situ hybridization study of fatal cases , 2004, The Journal of pathology.