Non-typeable Haemophilus influenzae biofilm formation on air-liquid interface cultured primary human airway epithelia

Organotypic cultures of primary human airway epithelial cells have been used to investigate the morphology, ion and fluid transport, innate immunity, transcytosis, infection, inflammation, signaling, cilia, and repair functions of this complex tissue. However, we do not know how closely these cultures resemble the airway surface epithelium in vivo. In this study, we examined the genome-wide expression profile of tracheal and bronchial human airway epithelia in vivo and compared it with the expression profile of primary cultures of human airway epithelia grown at the air-liquid interface. For comparison, we also investigated the expression profile of Calu-3 cells grown at the air-liquid interface and primary cultures of human airway epithelia submerged in nutrient media. We found that the transcriptional profile of differentiated primary cultures grown at the air-liquid interface most closely resembles that of in vivo airway epithelia, suggesting that the use of primary cultures and the presence of an air-liquid interface are important to recapitulate airway epithelia biology. We describe a high level of similarity between cells of tracheal and bronchial origin within and between different human donors, which suggests a very robust expression profile that is specific to airway cells.

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A simple method for testing the toxicity of nanomaterials on 3D air–liquid interface human airway epithelia (MucilAir™)

Toxicology Letters ◽

10.1016/j.toxlet.2011.05.962 ◽

2011 ◽

Vol 205 ◽

pp. S284 ◽

Cited By ~ 1

Author(s):

S. Constant ◽

S. Huang ◽

M. Caulfuty ◽

R. Bonfante ◽

M. Monachino ◽

...

Keyword(s):

Liquid Interface ◽

Simple Method ◽

Airway Epithelia ◽

Human Airway ◽

Air Liquid Interface ◽

Human Airway Epithelia

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Biofilm formation on human airway epithelia by encapsulated Neisseria meningitidis serogroup B

Microbes and Infection ◽

10.1016/j.micinf.2008.12.001 ◽

2009 ◽

Vol 11 (2) ◽

pp. 281-287 ◽

Cited By ~ 21

Author(s):

R. Brock Neil ◽

Jian Q. Shao ◽

Michael A. Apicella

Keyword(s):

Neisseria Meningitidis ◽

Biofilm Formation ◽

Airway Epithelia ◽

Human Airway ◽

Human Airway Epithelia

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Role of Actin Filament Network in Burkholderia multivorans Invasion in Well-Differentiated Human Airway Epithelia

Infection and Immunity ◽

10.1128/iai.71.11.6607-6609.2003 ◽

2003 ◽

Vol 71 (11) ◽

pp. 6607-6609 ◽

Cited By ~ 11

Author(s):

Ute E. Schwab ◽

Carla M. P. Ribeiro ◽

Heiner Neubauer ◽

Richard C. Boucher

Keyword(s):

Biofilm Formation ◽

Cell Entry ◽

Airway Epithelia ◽

Burkholderia Multivorans ◽

Human Airway ◽

Internalization Process ◽

Well Differentiated ◽

Filament Network ◽

Human Airway Epithelia

ABSTRACT The role of the actin-based cytoskeleton in the internalization process of Burkholderia multivorans by well-differentiated human airway epithelia was investigated by immunohistology and confocal microscopy. Our data suggest that an intact actin cytoskeleton is required for biofilm formation but not single cell entry or paracytosis.

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The SARS-CoV-2 Transcriptome and the Dynamics of the S Gene Furin Cleavage Site in Primary Human Airway Epithelia

mBio ◽

10.1128/mbio.01006-21 ◽

2021 ◽

Vol 12 (3) ◽

Author(s):

Wei Zou ◽

Min Xiong ◽

Siyuan Hao ◽

Elizabeth Yan Zhang ◽

Nathalie Baumlin ◽

...

Keyword(s):

Cleavage Site ◽

Vero Cells ◽

High Rate ◽

Furin Cleavage ◽

Airway Epithelia ◽

Human Airway ◽

Furin Cleavage Site ◽

Air Liquid Interface ◽

Human Airway Epithelia

ABSTRACT The spike (S) polypeptide of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) consists of the S1 and S2 subunits and is processed by cellular proteases at the S1/S2 boundary that contains a furin cleavage site (FCS), 682RRAR↓S686. Various deletions surrounding the FCS have been identified in patients. When SARS-CoV-2 propagated in Vero cells, it acquired deletions surrounding the FCS. We studied the viral transcriptome in Vero cell-derived SARS-CoV-2-infected primary human airway epithelia (HAE) cultured at an air-liquid interface (ALI) with an emphasis on the viral genome stability of the FCS. While we found overall the viral transcriptome is similar to that generated from infected Vero cells, we identified a high percentage of mutated viral genome and transcripts in HAE-ALI. Two highly frequent deletions were found at the FCS region: a 12 amino acid deletion (678TNSPRRAR↓SVAS689) that contains the underlined FCS and a 5 amino acid deletion (675QTQTN679) that is two amino acids upstream of the FCS. Further studies on the dynamics of the FCS deletions in apically released virions from 11 infected HAE-ALI cultures of both healthy and lung disease donors revealed that the selective pressure for the FCS maintains the FCS stably in 9 HAE-ALI cultures but with 2 exceptions, in which the FCS deletions are retained at a high rate of >40% after infection of ≥13 days. Our study presents evidence for the role of unique properties of human airway epithelia in the dynamics of the FCS region during infection of human airways, which is likely donor dependent. IMPORTANCE Polarized human airway epithelia at an air-liquid interface (HAE-ALI) are an in vitro model that supports efficient infection of SARS-CoV-2. The spike (S) protein of SARS-CoV-2 contains a furin cleavage site (FCS) at the boundary of the S1 and S2 domains which distinguishes it from SARS-CoV. However, FCS deletion mutants have been identified in patients and in vitro cell cultures, and how the airway epithelial cells maintain the unique FCS remains unknown. We found that HAE-ALI cultures were capable of suppressing two prevalent FCS deletion mutants (Δ678TNSPRRAR↓SVAS689 and Δ675QTQTN679) that were selected during propagation in Vero cells. While such suppression was observed in 9 out of 11 of the tested HAE-ALI cultures derived from independent donors, 2 exceptions that retained a high rate of FCS deletions were also found. Our results present evidence of the donor-dependent properties of human airway epithelia in the evolution of the FCS during infection.

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