A novel in vitro model of primary human pediatric lung epithelial cells

Wang, Qian; Bhattacharya, Soumyaroop; Mereness, Jared A.; Anderson, Christopher; Lillis, Jacquelyn; Misra, Ravi S.; Romas, Stephen; Huyck, Heidie; Howell, A.; Bandyopadhyay, Gautam; Donlon, Kathy; Myers, Jason; Ashton, John M.; Pryhuber, Gloria S.; Mariani, Thomas J.

doi:10.1038/s41390-019-0340-9

Cited by 22 publications

(30 citation statements)

References 35 publications

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“…Primary pediatric human lung epithelial (PHLE) cells were isolated from distal lung tissue digests of donor-quality normal lung tissue from 1-dayeold, 2-montheold, and 9-montheold donors, which were obtained by the human tissue core of the Molecular Atlas of Lung Development Program at the University of Rochester. 42,43 Lung tissue was digested with a protease cocktail containing collagenase A (101033578001; Sigma-Aldrich, St. Louis, MO), dispase (354235; Corning, Corning, NY), elastase (LS002290; Worthington-Biochem, Lakewood, NJ), and DNAase (DN25; Sigma-Aldrich). Dissociated cells were washed twice in Dulbecco's PBS containing 1% penicillin-streptomycin (15140122; Gibco), 50 mg/ mL gentamicin, and 0.25 mg/mL amphotericin B and centrifuged with 800 Â g for 10 minutes.…”

Section: Cell Culturementioning

confidence: 99%

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Collagen VI Deficiency Results in Structural Abnormalities in the Mouse Lung

Mereness

Bhattacharya

Ren

et al. 2020

The American Journal of Pathology

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Collagen VI (COL6) is known for its role in a spectrum of congenital muscular dystrophies, which are often accompanied by respiratory dysfunction. However, little is known regarding the function of COL6 in the lung. We confirmed the presence of COL6 throughout the basement membrane region of mouse lung tissue. Lung structure and organization were studied in a previously described Col6a1 À/À mouse, which does not produce detectable COL6 in the lung. The Col6a1 À/À mouse displayed histopathologic alveolar and airway abnormalities. The airspaces of Col6a1 À/À lungs appeared simplified, with larger (29%; P < 0.01) and fewer (31%; P < 0.001) alveoli. These airspace abnormalities included reduced isolectin B4 þ alveolar capillaries and surfactant protein Cepositive alveolar epithelial type-II cells. Alterations in lung function consistent with these histopathologic changes were evident. Col6a1 À/À mice also displayed multiple airway changes, including increased branching (59%; P < 0.001), increased mucosal thickness (34%; P < 0.001), and increased epithelial cell density (13%; P < 0.001). Comprehensive transcriptome analysis revealed that the loss of COL6 is associated with reductions in integrin-paxillinephosphatidylinositol 3-kinase signaling in vivo. In vitro, COL6 promoted steady-state phosphorylated paxillin levels and reduced cell density (16% to 28%; P < 0.05) at confluence. Inhibition of phosphatidylinositol 3-kinase, or its downstream effectors, resulted in increased cell density to a level similar to that seen on matrices lacking COL6. (Am J Pathol 2020, 190: 426e441; https:// The extracellular matrix (ECM) is composed of the noncellular components of a tissue and provides multiple critical structural and signaling properties. Major components of the ECM include collagens, proteoglycans, glycoproteins, elastin, and fibronectin. Highly diverse organization of the ECM contributes a broad range of properties, including tissue stiffness and elasticity, to facilitate proper tissue and organ function. 1e3 In addition to structural properties, ECM can provide numerous signaling cues for events, including migration, proliferation, and differentiation, to direct development, healing, and homeostatic conditions. 4,5 This can be accomplished via direct interaction of cells with the matrix or by sequestration and release or presentation of growth factors by the matrix. Disruption of any of the functions of the ECM can be devastating to the tissue and is known to be involved in a multitude of diseases. 6The ECM is known to be critical in the development, organization, and maintenance of lung structure. During development and homeostasis, organization and presence of particular ECM components direct both cellular function and further remodeling and maturation of the matrix by the surrounding cells. This can be due to cell signaling initiated by interaction with matrix components or mechanotransduction, a process by which cells respond to the mechanical forces

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Section: Cell Culturementioning

confidence: 99%

“…These expression data suggest that PHLE cells most closely resemble distal airway epithelium. 43 All experiments were conducted using PHLE cells between passages 1 and 3.…”

Section: Cell Culturementioning

confidence: 99%

Collagen VI Deficiency Results in Structural Abnormalities in the Mouse Lung

Mereness

Bhattacharya

Ren

et al. 2020

The American Journal of Pathology

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“…Pediatric/Infant Human Lung Epithelial (PHLE) cells were generated from transplant donor quality lung from the LungMAP program as we recently described 45…”

Section: Development Of In Vitro Rsv Infection Model Using Respiratormentioning

confidence: 99%

Insufficiency in airway interferon activation defines clinical severity to infant RSV infection

Chu

Qiu

McCall

et al. 2019

Preprint

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Respiratory syncytial virus (RSV) is the leading cause of severe respiratory disease in infants. Other than age at the time of infection, the causes and correlates of severe illness in infants lacking known risk factors are poorly defined. We recruited a cohort of confirmed RSV-infected infants and simultaneously assayed the presence of resident airway microbiota and the molecular status of their airways using a novel method. Rigorous statistical analyses identified a molecular airway gene expression signature of severe illness dominated by excessive chemokine expression. Global 16S rRNA sequencing confirmed an association between H. influenzae and clinical severity. Interestingly, adjusting for H. influenzae in our gene expression analysis revealed an association between severity and airway lymphocyte accumulation. Exploring the relationship between airway gene expression and the time of onset of clinical symptoms revealed a robust, acute activation of interferon (IFN) signaling, which was absent in subjects with severe illness. Finally, we explored the relationship between IFN activity, airway gene expression and productive RSV infection using a novel in vitro model of bona fide pediatric human airway epithelial cells. Interestingly, blocking IFN signaling, but not IFN ligand production, in these cells leads to increased viral infection. Our data reveal that acute airway interferon responses are physiologically relevant in the context of infant RSV infection and may be a target for therapeutic intervention. Additionally, the airway gene expression signature we define may be useful as a biomarker for efficacy of intervention responses. All rights reserved. No reuse allowed without permission.

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“…In addition, lung epithelial cells can produce a variety of inflammatory cytokines (e.g., that can influence the course of T cell responses (19). Lung epithelial cells in infants differ from those in adults in humans (20)(21)(22)(23) and animals (24). The potential effect of lung epithelial cells in the immune responses of infants is a recent area of inquiry, and could be a significant determining factor in their apparent altered immune response during influenza virus infection.…”

Section: Introductionmentioning

confidence: 99%

Protective Intranasal Immunization Against Influenza Virus in Infant Mice Is Dependent on IL-6

et al. 2020

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Respiratory diseases adversely affect infants and are the focus of efforts to develop vaccinations and other modalities to prevent disease. The infant immune system differs from that of older children and adults in many ways that are as yet ill understood. We have used a C57BL/6 mouse model of infection with a laboratory-adapted strain of influenza (PR8) to delineate the importance of the cytokine IL-6 in the innate response to primary infection and in the development of protective immunity in adult mice. Herein, we used this same model in infant (14 days of age) mice to determine the effect of IL-6 deficiency. Infant wild type mice are more susceptible than older mice to infection, similar to the findings in humans. IL-6 is expressed in the lung in the early response to PR8 infection. While intramuscular immunization does not protect against lethal challenge, intranasal administration of heat inactivated virus is protective and correlates with expression of IL-6 in the lung, activation of lung CD8 cells, and development of an influenza-specific antibody response. In IL-6 deficient mice, this response is abrogated, and deficient mice are not protected against lethal challenge. These studies support the importance of the role of the tissue environment in infant immunity, and further suggest that IL-6 may be helpful in the generation of protective immune responses in infants.

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A novel in vitro model of primary human pediatric lung epithelial cells

Cited by 22 publications

References 35 publications

Collagen VI Deficiency Results in Structural Abnormalities in the Mouse Lung

Collagen VI Deficiency Results in Structural Abnormalities in the Mouse Lung

Insufficiency in airway interferon activation defines clinical severity to infant RSV infection

Protective Intranasal Immunization Against Influenza Virus in Infant Mice Is Dependent on IL-6

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