scholarly journals Infection of mice with influenza A/WSN/33 (H1N1) virus alters alveolar type II cell phenotype

2015 ◽  
Vol 308 (7) ◽  
pp. L628-L638 ◽  
Author(s):  
Christian C. Hofer ◽  
Parker S. Woods ◽  
Ian C. Davis
Keyword(s):  
Influenza A ◽  
Influenza Infection ◽  
Influenza Viruses ◽  
Normal Lung ◽  
Cell Phenotype ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii ◽  
Atii Cell ◽  
The Impact

Influenza viruses cause acute respiratory disease of great importance to public health. Alveolar type II (ATII) respiratory epithelial cells are central to normal lung function and are a site of influenza A virus replication in the distal lung. However, the consequences of infection for ATII cell function are poorly understood. To determine the impact of influenza infection on ATII cells we used C57BL/6-congenic SP-CGFP mice that express green fluorescent protein (GFP) under the control of the surfactant protein-C (SP-C) promoter, which is only active in ATII cells. Most cells isolated from the lungs of uninfected SP-CGFP mice were GFP+ but did not express the alveolar type I (ATI) antigen podoplanin (PODO). ATII cells were also EpCAM+ and α2,3-linked sialosaccharide+. Infection with influenza A/WSN/33 virus caused severe hypoxemia and pulmonary edema. This was accompanied by loss of whole lung GFP fluorescence, reduced ATII cell yields, increased ATII cell apoptosis, reduced SP-C gene and protein expression in ATII cell lysates, and increased PODO gene and protein levels. Flow cytometry indicated that infection decreased GFP+/PODO− cells and increased GFP−/PODO+ and GFP−/PODO− cells. Very few GFP+/PODO+ cells were detectable. Finally, infection resulted in a significant decline in EpCAM expression by PODO+ cells, but had limited effects on α2,3-linked sialosaccharides. Our findings indicate that influenza infection results in a progressive differentiation of ATII cells into ATI-like cells, possibly via an SP-C−/PODO− intermediate, to replace dying or dead ATI cells. However, impaired SP-C synthesis is likely to contribute significantly to reduced lung compliance in infected mice.

Modulation of pulmonary alveolar type II cell phenotype and communication by extracellular matrix and KGF

2001 ◽  
Vol 281 (4) ◽  
pp. C1291-C1299 ◽  
Author(s):  
Brant E. Isakson ◽  
Richard L. Lubman ◽  
Gregory J. Seedorf ◽  
Scott Boitano
Keyword(s):  
Extracellular Matrix ◽  
Growth Factor ◽  
Type I Collagen ◽  
Cultured Cells ◽  
Cell Phenotype ◽  
Alveolar Type ◽  
Specific Cell ◽  
Specific Marker ◽  
Type I ◽  
Type Ii

The alveolar epithelium consists of two cell types, alveolar type I (AT1) and alveolar type II (AT2) cells. We have recently shown that 7-day-old cultures of AT2 cells grown on a type I collagen/fibronectin matrix develop phenotypic characteristics of AT1 cells, display a distinct connexin profile, and coordinate mechanically induced intercellular Ca2+ changes via gap junctions (25). In this study, we cultured AT2 cells for 7 days on matrix supplemented with laminin-5 and/or in the presence of keratinocyte growth factor. Under these conditions, cultured AT2 cells display AT2 type morphology, express the AT2-specific marker surfactant protein C, and do not express AT1-specific cell marker aquaporin 5, all consistent with maintenance of AT2 phenotype. These AT2-like cells also coordinate mechanically induced intercellular Ca2+ signaling, but, unlike AT1-like cells, do so by using extracellular nucleotide triphosphate release. Additionally, cultured cells that retain AT2 cell-specific markers express connexin profiles different from cultured cells with AT1 characteristics. The parallel changes in intercellular Ca2+ signaling with cell differentiation suggest that cell signaling mechanisms are an intrinsic component of lung alveolar cell phenotype. Because lung epithelial injury is accompanied by extracellular matrix and growth factor changes, followed by extensive cell division, differentiation, and migration of AT2 progenitor cells, we suggest that similar changes may be vital to the lung recovery and repair process in vivo.

scholarly journals Lethal H1N1 influenza A virus infection alters the murine alveolar type II cell surfactant lipidome

2016 ◽  
Vol 311 (6) ◽  
pp. L1160-L1169 ◽  
Author(s):  
Parker S. Woods ◽  
Lauren M. Doolittle ◽  
Lucia E. Rosas ◽  
Lisa M. Joseph ◽  
Edward P. Calomeni ◽  
...  
Keyword(s):  
Acute Respiratory Distress Syndrome ◽  
Lipid Metabolism ◽  
Influenza A ◽  
Bronchoalveolar Lavage Fluid ◽  
Distress Syndrome ◽  
Lavage Fluid ◽  
Alveolar Type ◽  
Type Ii ◽  
Atii Cell ◽  
Surfactant Lipid

Alveolar type II (ATII) epithelial cells are the primary site of influenza virus replication in the distal lung. Development of acute respiratory distress syndrome in influenza-infected mice correlates with significant alterations in ATII cell function. However, the impact of infection on ATII cell surfactant lipid metabolism has not been explored. C57BL/6 mice were inoculated intranasally with influenza A/WSN/33 (H1N1) virus (10,000 plaque-forming units/mouse) or mock-infected with virus diluent. ATII cells were isolated by a standard lung digestion protocol at 2 and 6 days postinfection. Levels of 77 surfactant lipid-related compounds of known identity in each ATII cell sample were measured by ultra-high-performance liquid chromatography-mass spectrometry. In other mice, bronchoalveolar lavage fluid was collected to measure lipid and protein content using commercial assay kits. Relative to mock-infected animals, ATII cells from influenza-infected mice contained reduced levels of major surfactant phospholipids (phosphatidylcholine, phosphatidylglycerol, and phosphatidylethanolamine) but increased levels of minor phospholipids (phosphatidylserine, phosphatidylinositol, and sphingomyelin), cholesterol, and diacylglycerol. These changes were accompanied by reductions in cytidine 5′-diphosphocholine and 5′-diphosphoethanolamine (liponucleotide precursors for ATII cell phosphatidylcholine and phosphatidylethanolamine synthesis, respectively). ATII cell lamellar bodies were ultrastructurally abnormal after infection. Changes in ATII cell phospholipids were reflected in the composition of bronchoalveolar lavage fluid, which contained reduced amounts of phosphatidylcholine and phosphatidylglycerol but increased amounts of sphingomyelin, cholesterol, and protein. Influenza infection significantly alters ATII cell surfactant lipid metabolism, which may contribute to surfactant dysfunction and development of acute respiratory distress syndrome in influenza-infected mice.

scholarly journals The Rho Pathway Mediates Transition to an Alveolar Type I Cell Phenotype During Static Stretch of Alveolar Type II Cells

2010 ◽  
Vol 67 (6) ◽  
pp. 585-590 ◽  
Author(s):  
Cherie D Foster ◽  
Linda S Varghese ◽  
Linda W Gonzales ◽  
Susan S Margulies ◽  
Susan H Guttentag
Keyword(s):  
Cell Phenotype ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii Cells ◽  
Type Ii ◽  
Alveolar Type Ii Cells ◽  
Static Stretch ◽  
Alveolar Type I Cell

Freshly isolated rat alveolar type I cells, type II cells, and cultured type II cells have distinct molecular phenotypes

2005 ◽  
Vol 288 (1) ◽  
pp. L179-L189 ◽  
Author(s):  
Robert Gonzalez ◽  
Yee Hwa Yang ◽  
Chandi Griffin ◽  
Lennell Allen ◽  
Zachary Tigue ◽  
...  
Keyword(s):  
Gene Expression ◽  
Differentially Expressed Genes ◽  
Expression Profiles ◽  
Differentially Expressed ◽  
Cell Phenotype ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii Cells ◽  
Type Ii ◽  
Isolated Rat

We used microarray analysis with Affymetrix rat chips to determine gene expression profiles of freshly isolated rat type I (TI) and TII cells and cultured TII cells. Our goals were 1) to describe molecular phenotypic “fingerprints” of TI and TII cells, 2) to gain insight into possible functional differences between the two cell types through differentially expressed genes, 3) to identify genes that might indicate potential functions of TI cells, since so little is known about this cell type, and 4) to ascertain the similarities and differences in gene expression between cultured TII cells and freshly isolated TI cells. For these experiments, we used preparations of isolated TI and TII cells that contained <2% cross-contamination. With a false discovery rate of 1%, 601 genes demonstrated over twofold different expression between TI and TII cells. Those genes with very high levels of differential expression may be useful as markers of cell phenotype and in generating novel hypotheses about functions of TI and TII cells. We found similar numbers of differentially expressed genes between freshly isolated TI or TII cells and cultured TII cells (698, 637 genes) and freshly isolated TI and TII cells (601 genes). Tests of sameness/difference including cluster dendrograms and log/log identity plots indicated major differences between the phenotypes of freshly isolated TI cell and cultured type II cell populations. The latter results suggest that experiments with TII cells cultured under these conditions should be interpreted with caution with respect to biological relevance to TI or TII cells.

scholarly journals 2750. Sequential Influenza A H1N1 and Influenza A H3N2 Challenge Infections in Healthy Volunteers

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S969-S969 ◽  
Author(s):  
Alison Han ◽  
Luca Giurgea ◽  
Adriana Cervantes-Medina ◽  
Kristina Edwards ◽  
Luz Angela Rosas ◽  
...  
Keyword(s):  
Healthy Volunteers ◽  
Influenza A ◽  
Reverse Genetics ◽  
Clinical Symptoms ◽  
Influenza Infection ◽  
Influenza Viruses ◽  
Wild Type ◽  
Viral Shedding ◽  
Influenza A H1n1 ◽  
The Impact

Abstract Background Seasonal influenza causes significant annual morbidity and mortality. The effects of yearly exposures on immunity are not clear and recent observations have demonstrated that long lasting protection against a matched strain may not naturally occur. The 2018–2019 influenza season consisted of an initial peak of H1N1 infections followed by a wave of H3N2 infections. These consecutive waves raise questions about how influenza immunity is affected by sequential exposure to different influenza strains. Challenge studies provide a unique opportunity to study this phenomenon. Here we describe a subset of participants who were sequentially infected in two separate challenge studies with wild-type H1N1 and H3N2 viruses. Methods Healthy volunteers completed two sequential influenza challenge studies at the NIH Clinical Center. Participants were inoculated with reverse genetics, cell-based, GMP wild-type influenza viruses, A(H1N1)pdm09 and A(H3N2) strains. Participants remained isolated in the hospital for a minimum of 9 days and were monitored daily for viral shedding and clinical symptoms. After discharge, participants were followed for 2 months. Results Between 2014 and 2017, 14 healthy volunteers were exposed to Influenza A(H1N1) and Influenza A(H3N2). Time between infections ranged from 2 months to 2 years. Thirteen (93%) participants developed confirmed influenza infection after H1N1 challenge and 9 (64%) after H3N2 challenge. Eight (57%) participants developed confirmed infections after both exposures. Variable degrees of symptoms, shedding, and disease severity were observed. Systemic antibody responses to the HA and NA of both H1N1 and H3N2 varied over time during these sequential infections. Conclusion More than half of all participants who completed 2 sequential H1N1 and H3N2 challenge studies demonstrated confirmed infection to both viruses. These sequential infections had varying effects on the disease experienced and the immunity that developed after infection. These observations are important in understanding the impact of sequential exposures on influenza immunity. Disclosures All authors: No reported disclosures.

scholarly journals Characteristics of Cl− uptake in rat alveolar type I cells

2009 ◽  
Vol 297 (5) ◽  
pp. L816-L827 ◽  
Author(s):  
Meshell Johnson ◽  
Lennell Allen ◽  
Leland Dobbs
Keyword(s):  
Fetal Lung ◽  
Fluid Secretion ◽  
Normal Lung ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii Cells ◽  
Type Ii ◽  
Adult Lung ◽  
Type I Cells ◽  
I Cells

Although Cl− transport in fetal lung is important for fluid secretion and normal lung development, the role of Cl− transport in adult lung is not well understood. In physiological studies, the cystic fibrosis transmembrane regulator (CFTR) plays a role in fluid absorption in the distal air spaces of adult lung, and alveolar type II cells cultured for 5 days have the capacity to transport Cl−. Although both alveolar type I and type II cells express CFTR, it has previously not been known whether type I cells transport Cl−. We studied Cl− uptake in isolated type I cells directly, using either radioisotopic tracers or halide-sensitive fluorescent indicators. By both methods, type I cells take up Cl−. In the presence of β-adrenergic agonist stimulation, Cl− uptake can be inhibited by CFTR antagonists. Type I cells express both the Cl−/HCO3− anion exchanger AE2 and the voltage-gated Cl− channels CLC5 and CLC2. Inhibitors of AE2 also block Cl− uptake in type I cells. Together, these results demonstrate that type I cells are capable of Cl− uptake and suggest that the effects seen in whole lung studies establishing the importance of Cl− movement in alveolar fluid clearance may be, in part, the result of Cl− transport across type I cells.

scholarly journals Developing a system to estimate the severity of influenza infection in England: findings from a hospital-based surveillance system between 2010/2011 and 2014/2015

2017 ◽  
Vol 145 (7) ◽  
pp. 1461-1470 ◽  
Author(s):  
N. L. BODDINGTON ◽  
N. Q. VERLANDER ◽  
R. G. PEBODY
Keyword(s):  
Surveillance System ◽  
Influenza A ◽  
Influenza Pandemic ◽  
Influenza Infection ◽  
Rapid Assessment ◽  
Age Groups ◽  
Influenza Viruses ◽  
Influenza Surveillance ◽  
High Dependency ◽  
The Impact

SUMMARYThe UK Severe Influenza Surveillance System (USISS) was established following the 2009 influenza pandemic to monitor severe seasonal influenza. This article describes the severity of influenza observed in five post-2009 pandemic seasons in England. Two key measures were used to assess severity: impact measured through the cumulative incidence of laboratory-confirmed hospitalised influenza and case severity through the proportion of confirmed hospitalised cases admitted into intensive care units (ICU)/high dependency units (HDU). The impact of influenza varied by subtype and age group across the five seasons with the highest crude cumulative hospitalisation incidence for influenza A/H1N1pdm09 cases in 2010/2011 and in 0–4 year olds each season for all-subtypes. Case severity also varied by subtype and season with a higher hospitalisation: ICU ratio for A/H1N1pdm09 and older age groups (older than 45 years). The USISS system provides a tool for measuring severity of influenza each year. Such seasonal surveillance can provide robust baseline estimates to allow for rapid assessment of the severity of seasonal and emerging influenza viruses.

scholarly journals LAMP3 is critical for surfactant homeostasis in mice

2021 ◽  
Author(s):  
Lars P. Lunding ◽  
Daniel Krause ◽  
Guido Stichtenoth ◽  
Cordula Stamme ◽  
Niklas Lauterbach ◽  
...  
Keyword(s):  
Lung Function ◽  
Lipid Composition ◽  
Knockout Mice ◽  
Pulmonary Surfactant ◽  
Normal Lung ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii ◽  
Average Life Span ◽  
Deficient Mice

AbstractLysosome-associated membrane glycoprotein 3 (LAMP3) is a type I transmembrane protein of the LAMP protein family with a cell-type-specific expression in alveolar type II cells in mice and hitherto unknown function. In type II pneumocytes, LAMP3 is localized in lamellar bodies, secretory organelles releasing pulmonary surfactant into the extracellular space to lower surface tension at the air/liquid interface. The physiological function of LAMP3, however, remains enigmatic. We generated Lamp3 knockout mice by CRISPR/Cas9. LAMP3 deficient mice are viable with an average life span and display regular lung function under basal conditions. The levels of a major hydrophobic protein component of pulmonary surfactant, SP-C, are strongly increased in the lung of Lamp3 knockout mice, and the lipid composition of the bronchoalveolar lavage shows mild but significant changes, resulting in alterations in surfactant functionality. In ovalbumin-induced experimental allergic asthma, the changes in lipid composition are aggravated, and LAMP3-deficient mice exert an increased airway resistance. Our data suggest a critical role of LAMP3 in the regulation of pulmonary surfactant homeostasis and normal lung function.

scholarly journals LAMP3 deficiency affects surfactant homeostasis in mice

PLoS Genetics ◽  
2021 ◽  
Vol 17 (6) ◽  
pp. e1009619
Author(s):  
Lars P. Lunding ◽  
Daniel Krause ◽  
Guido Stichtenoth ◽  
Cordula Stamme ◽  
Niklas Lauterbach ◽  
...  
Keyword(s):  
Lung Function ◽  
Lipid Composition ◽  
Knockout Mice ◽  
Pulmonary Surfactant ◽  
Normal Lung ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii ◽  
Average Life Span ◽  
Deficient Mice

Lysosome-associated membrane glycoprotein 3 (LAMP3) is a type I transmembrane protein of the LAMP protein family with a cell-type-specific expression in alveolar type II cells in mice and hitherto unknown function. In type II pneumocytes, LAMP3 is localized in lamellar bodies, secretory organelles releasing pulmonary surfactant into the extracellular space to lower surface tension at the air/liquid interface. The physiological function of LAMP3, however, remains enigmatic. We generated Lamp3 knockout mice by CRISPR/Cas9. LAMP3 deficient mice are viable with an average life span and display regular lung function under basal conditions. The levels of a major hydrophobic protein component of pulmonary surfactant, SP-C, are strongly increased in the lung of Lamp3 knockout mice, and the lipid composition of the bronchoalveolar lavage shows mild but significant changes, resulting in alterations in surfactant functionality. In ovalbumin-induced experimental allergic asthma, the changes in lipid composition are aggravated, and LAMP3-deficient mice exert an increased airway resistance. Our data suggest a critical role of LAMP3 in the regulation of pulmonary surfactant homeostasis and normal lung function.

Invited Review: Biophysical properties of sodium channels in lung alveolar epithelial cells

2002 ◽  
Vol 93 (5) ◽  
pp. 1852-1859 ◽  
Author(s):  
Sadis Matalon ◽  
Ahmed Lazrak ◽  
Lucky Jain ◽  
Douglas C. Eaton
Keyword(s):  
Sodium Channels ◽  
Sodium Transport ◽  
Normal Lung ◽  
Epithelial Tissue ◽  
Alveolar Type ◽  
Type I ◽  
Type Ii Cells ◽  
Type Ii ◽  
Biophysical Properties ◽  
Alveolar Type Ii Cells

Amiloride-sensitive sodium channels in the lung play an important role in lung fluid balance. Particularly in the alveoli, sodium transport is closely regulated to maintain an appropriate fluid layer on the surface of the alveoli. Alveolar type II cells appear to play an important role in this sodium transport, with the role of alveolar type I cells being less clear. In alveolar type II cells, there are a variety of different amiloride-sensitive, sodium-permeable channels. This significant diversity appears to play a role in both normal lung physiology and in pathological states. In many epithelial tissues, amiloride-sensitive epithelial sodium channels (ENaC) are formed from three subunit proteins, designated α-, β-, and γ-ENaC. At least part of the diversity of sodium-permeable channels in lung arises from the assembling of different combinations of these subunits to form channels with different biophysical properties and different mechanisms for regulation. This leads to epithelial tissue in the lung, which has enormous flexibility to alter the magnitude and regulation of salt and water transport. In this review, we discuss the biophysical properties and occurrence of these various channels and some of the mechanisms for their regulation.

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