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

Mossel, Eric C.; Wang, Jieru; Jeffers, Scott A.; Edeen, Karen E.; Wang, Shuanglin; Cosgrove, Gregory P.; Funk, C. Joel; Manzer, Rizwan; Miura, Tanya A.; Pearson, Leonard; Holmes, Kathryn V.; Mason, Robert J.

doi:10.1016/j.virol.2007.09.045

Cited by 158 publications

(149 citation statements)

References 40 publications

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“…In our previous study (18), we detected no SARS-CoV infection of human alveolar type I-like cells under submerged conditions, and ACE2 protein expression was below the level of detection. In pilot experiments with unselected donors, we detected very little infection of type I-like cells under A/L conditions at 24 hours after inoculation.…”

Section: Some Alveolar Type I-like Cells Are Susceptible To Sars-cov mentioning

confidence: 76%

“…In our earlier study with submerged human alveolar type II cells on inserts, we detected a very low percentage of infected cells and could not evaluate their innate immune response (18). However, culturing and infecting the type II cell at an air-liquid interface (A/L) allowed for significant infection and characterization of their innate immune response.…”

Section: Clinical Relevancementioning

confidence: 96%

“…To test whether type II cells cultured at the A/L were more susceptible to SARS-CoV infection than type II cells in submerged cultures (18), human type II cells cultured at an A/L on six-well inserts were inoculated on the apical surface with 250 ml of medium containing SARS-CoV at an estimated multiplicity of 2 pfu/ cell, and allowed to adsorb for 4 hours at 37 8 C. Then the apical fluid and inoculum were aspirated, and cells were maintained under A/L conditions. SARS-CoV reproducibly caused productive infection of the primary human alveolar type II cells cultured at the A/L.…”

Section: Sars-cov Readily Infects Primary Human Alveolar Type II Cellmentioning

confidence: 99%

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Innate Immune Response of Human Alveolar Type II Cells Infected with Severe Acute Respiratory Syndrome–Coronavirus

Qian

Travanty

Oko

et al. 2013

Am J Respir Cell Mol Biol

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286

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Severe acute respiratory syndrome (SARS)-coronavirus (CoV) produces a devastating primary viral pneumonia with diffuse alveolar damage and a marked increase in circulating cytokines. One of the major cell types to be infected is the alveolar type II cell. However, the innate immune response of primary human alveolar epithelial cells infected with SARS-CoV has not been defined. Our objectives included developing a culture system permissive for SARS-CoV infection in primary human type II cells and defining their innate immune response. Culturing primary human alveolar type II cells at an airliquid interface (A/L) improved their differentiation and greatly increased their susceptibility to infection, allowing us to define their primary interferon and chemokine responses. Viral antigens were detected in the cytoplasm of infected type II cells, electron micrographs demonstrated secretory vesicles filled with virions, virus RNA concentrations increased with time, and infectious virions were released by exocytosis from the apical surface of polarized type II cells. A marked increase was evident in the mRNA concentrations of interferon-b and interferon-l (IL-29) and in a large number of proinflammatory cytokines and chemokines. A surprising finding involved the variability of expression of angiotensin-converting enzyme-2, the SARS-CoV receptor, in type II cells from different donors. In conclusion, the cultivation of alveolar type II cells at an air-liquid interface provides primary cultures in which to study the pulmonary innate immune responses to infection with SARS-CoV, and to explore possible therapeutic approaches to modulating these innate immune responses.Keywords: lung innate immune response; cytokine responses to SARS coronavirus; lung cell differentiation; air-liquid interface culturesSevere acute respiratory syndrome-associated coronavirus (SARSCoV) produces devastating viral pneumonia (1, 2). Pathologic changes are most prominent in the lungs, with disruptions of the epithelium in gas exchange areas and conducting airways (2-4). The epithelial cells of the alveoli and the conducting airways are the primary targets of SARS-CoV in the human lung, and they express the SARS receptor, angiotension-converting enzyme-2 (ACE2) (5-8). In autopsies of patients with SARS, coronavirus RNA and proteins have been detected in type II cells via immunocytochemisty and in situ hybridization (7,(9)(10)(11)(12)(13)(14). In the aged macaque model of SARS, in which the initial site of infection in the lung can be studied, virus infection was detected in both alveolar type I and type II cells (11,15). In human SARS autopsy specimens, the infection of alveolar macrophages was also suggested because they contained SARS antigens and formed multinucleated giant cells (2, 11). However, in vitro human alveolar macrophages, monocyte-derived dendritic cells, and monocytes are not readily susceptible to SARS-CoV, and these cell types show only very modest cytokine and interferon responses upon exposure to SARS-CoV (16-21).One of the major...

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Section: Some Alveolar Type I-like Cells Are Susceptible To Sars-cov mentioning

confidence: 76%

Section: Clinical Relevancementioning

confidence: 96%

Section: Sars-cov Readily Infects Primary Human Alveolar Type II Cellmentioning

confidence: 99%

See 1 more Smart Citation

Innate Immune Response of Human Alveolar Type II Cells Infected with Severe Acute Respiratory Syndrome–Coronavirus

Qian

Travanty

Oko

et al. 2013

Am J Respir Cell Mol Biol

Self Cite

286

266

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“…Infection of alveolar macrophages seemed to vary with the patient, with positive virus staining of CD68 cells in some individuals, but not others (Nicholls et al, 2006;Shieh et al, 2005;To et al, 2004). Using primary cell culture systems, SARS-CoV has been shown to replicate in type II cells, but not type Ilike cells and alveolar macrophages in vitro (Mossel et al, 2008). Human ciliated airway epithelial cell cultures and lung stem cells that express CCSP can also support growth of SARS-CoV (Ling et al, 2006;Sims et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Rat respiratory coronavirus infection: replication in airway and alveolar epithelial cells and the innate immune response

Funk

Manzer

Miura

et al. 2009

Journal of General Virology

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The rat coronavirus sialodacryoadenitis virus (SDAV) causes respiratory infection and provides a system for investigating respiratory coronaviruses in a natural host. A viral suspension in the form of a microspray aerosol was delivered by intratracheal instillation into the distal lung of 6-8-weekold Fischer 344 rats. SDAV inoculation produced a 7 % body weight loss over a 5 day period that was followed by recovery over the next 7 days. SDAV caused focal lesions in the lung, which were most severe on day 4 post-inoculation (p.i.). Immunofluorescent staining showed that four cell types supported SDAV virus replication in the lower respiratory tract, namely Clara cells, ciliated cells in the bronchial airway and alveolar type I and type II cells in the lung parenchyma. In bronchial alveolar lavage fluid (BALF) a neutrophil influx increased the population of neutrophils to 45 % compared with 6 % of the cells in control samples on day 2 after mock inoculation. Virus infection induced an increase in surfactant protein SP-D levels in BALF of infected rats on days 4 and 8 p.i. that subsided by day 12. The concentrations of chemokines MCP-1, LIX and CINC-1 in BALF increased on day 4 p.i., but returned to control levels by day 8. Intratracheal instillation of rats with SDAV coronavirus caused an acute, self-limited infection that is a useful model for studying the early events of the innate immune response to respiratory coronavirus infections in lungs of the natural virus host. INTRODUCTIONThe large volume of air inhaled each day and the extensive surface area of the lung makes the respiratory system especially vulnerable to airborne infectious agents. These pathogens include many respiratory viruses such as influenza virus, respiratory syncytial virus, rhinovirus and coronaviruses (CoVs). Respiratory tract infections are the leading cause of infectious disease globally (WHO, 2004). The outbreak of severe acute respiratory syndrome (SARS) in [2002][2003] emphasized the vulnerability of humans to respiratory virus diseases and the potential for high morbidity and mortality in viral infections of the lower respiratory tract. The aetiological agent of SARS was identified as a coronavirus (SARS-CoV) derived from an animal reservoir (Fouchier et al., 2003). Besides human infections, CoVs cause respiratory, enteric and neurological infections in a variety of birds and mammals (Weiss & Navas-Martin, 2005). Five human CoVs have been identified to date, all of which cause respiratory tract infections. Human CoVs 229E and OC43 were first identified in the 1960s, and they cause up to 15-35 % of the adult colds in otherwise healthy individuals (Kahn & McIntosh, 2005). After the discovery of SARS-CoV, two additional human CoVs were identified, NL63 and HKU1 (van der Hoek et al., 2004;Woo et al., 2005). On a global basis, the human CoVs 229E, OC43, HKU1 and NL63 are estimated to cause approximately 10 % of all hospitalizations of children for respiratory tract infections (Gerna et al., 2007;Vabret et al., 2008).A number of animal mod...

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“…In addition to participating in the formation of the air-blood barrier, they also transport water and ions, and exhibit certain immune regulatory functions (Mossel et al, 2008;Yu et al, 2011). These cells are distinct from type II alveolar cells, the involvement of which in lung injury has been comprehensively studied (Jamaluddin et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

Research Article Mac-1 deficiency induces respiratory failure by affecting type I alveolar epithelial cells.

Wang¹,

Ci²,

Liu³

et al. 2017

Genet. Mol. Res.

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ABSTRACT. As a b2 integrin family member, Mac-1 plays an important role in the inflammatory response. Inflammation and lung injury are closely associated, but the involvement of Mac-1 in the occurrence and development of such pathologies remains poorly understood. We aimed to investigate the relationship between Mac-1 deficiency and respiratory failure in Mac-1 knockout {Mac-1 -/-} mice, using C57BL/6J mice as a control. The newborn survival rate of Mac-1 -/-mice was calculated, and mouse lung tissue was treated with hematoxylin and eosin and subjected to immunofluorescent staining. Moreover, western blotting and immunohistochemistry were used to detect the expression of molecules specific to type I and type II alveolar epithelial cells, as well as alveolar surfactant proteins secreted by the latter. Survival of Mac-1 -/-pups was significantly lower than that of newborn C57BL/6J mice. In a float test, lung tissues from C57BL/6J mice were buoyant, whereas those of Mac-1 -/-mice were not. Compared with C57BL/6J mice, expression of proSP-C {specific to type II alveolar epithelial cells} and alveolar surfactant proteins in Mac-1 -/-mice was not significantly different, implying that type II cell function was unaltered. However, western blotting revealed expression of T1α, Aqp5, and Snx5 {type I alveolar epithelial cell markers} in Mac-1 -/-mice to be significantly decreased {P < 0.05}. In conclusion, Mac-1 may play an important role in respiratory failure. Its absence leads to this condition not by influencing type II alveolar epithelial cells or their secreted surfactant proteins, but rather by reducing type I alveolar cell numbers.

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

Cited by 158 publications

References 40 publications

Innate Immune Response of Human Alveolar Type II Cells Infected with Severe Acute Respiratory Syndrome–Coronavirus

Innate Immune Response of Human Alveolar Type II Cells Infected with Severe Acute Respiratory Syndrome–Coronavirus

Rat respiratory coronavirus infection: replication in airway and alveolar epithelial cells and the innate immune response

Research Article Mac-1 deficiency induces respiratory failure by affecting type I alveolar epithelial cells.

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