The effect of interleukin-13 (IL-13) and interferon-γ (IFN-γ) on expression of surfactant proteins in adult human alveolar type II cells in vitro

Background Pulmonary surfactant is a complex mixture of proteins and phospholipids synthetized by alveolar type II cells. Volatile anesthetics have been shown to reduce surfactant phospholipid biosynthesis by rat alveolar type II cells. Surfactant-associated protein C (SP-C) is critical for the alveolar surfactant functions. Our goal was to evaluate the effects of halothane and thiopental on SP-C messenger RNA (mRNA) expression in vitro in rat alveolar type II cells and in vivo in mechanically ventilated rats. Methods In vitro, freshly isolated alveolar type II cells were exposed to halothane during 4 h (1, 2, 4%) and 8 h (1%), and to thiopental during 4 h (10, 100 micrometer) and 8 h (100 micrometer). In vivo, rats were anesthetized with intraperitoneal thiopental or inhaled 1% halothane and mechanically ventilated for 4 or 8 h. SP-C mRNA expression was evaluated by ribonuclease protection assay. Results In vitro, 4-h exposure of alveolar type II cells to thiopental 10 and 100 micrometer increased their SP-C mRNA content to 145 and 197%, respectively, of the control values. In alveolar type II cells exposed for 4 h to halothane 1, 2, and 4%, the SP-C mRNA content increased dose-dependently to 160, 235, and 275%, respectively, of the control values. In vivo, in mechanically ventilated rats, 4 h of halothane anesthesia decreased the lung SP-C mRNA content to 53% of the value obtained in control (nonanesthetized, nonventilated) animals; thiopental anesthesia increased to 150% the lung SP-C mRNA content. Conclusions These findings indicate that halothane and thiopental used at clinically relevant concentrations modulate the pulmonary SP-C mRNA content in rats. In vivo, the additive role of mechanical ventilation is suggested.

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IL-17A And TNF-± Exert Synergistic Effects On Expression Of CXCL5 By Alveolar Type II Cells In Vivo And In Vitro

10.1164/ajrccm-conference.2010.181.1_meetingabstracts.a6787 ◽

2010 ◽

Author(s):

Yuhong Liu ◽

JunJie Mei ◽

Linda W. Gonzales ◽

Ping Wang ◽

Ning Dai ◽

...

Keyword(s):

Synergistic Effects ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells

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Effects of bone marrow-derived mesenchymal stromal cells on gene expression in human alveolar type II cells exposed to TNF-α , IL-1β , and IFN-γ

Physiological Reports ◽

10.14814/phy2.13831 ◽

2018 ◽

Vol 6 (16) ◽

pp. e13831 ◽

Cited By ~ 2

Author(s):

Matthew Schwede ◽

Erin M. Wilfong ◽

Rachel L. Zemans ◽

Patty J. Lee ◽

Claudia dos Santos ◽

...

Keyword(s):

Gene Expression ◽

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Tnf Α ◽

Ifn Γ

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Keratinocyte growth factor increases mRNAs for SP-A and SP-B in adult rat alveolar type II cells in culture

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1995.269.3.l344 ◽

1995 ◽

Vol 269 (3) ◽

pp. L344-L350 ◽

Cited By ~ 23

Author(s):

K. Sugahara ◽

J. S. Rubin ◽

R. J. Mason ◽

E. L. Aronsen ◽

J. M. Shannon

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Keratinocyte Growth Factor ◽

Primary Cultures ◽

Surfactant Proteins ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Adult Rat

The production of pulmonary surfactant, a complex of phospholipids and lung-specific surfactant proteins, is a primary function of alveolar type II cells. Although previous studies have demonstrated a role for cell-extracellular matrix interactions and normal cell shape in the maintenance of differentiated function in primary cultures of adult rat type II cells, a positive role for growth factors in surfactant protein gene expression in isolated normal adult type II cells has not been reported. In the present study, we have examined the effects of a panel of hormones, growth factors, and cytokines on the expression of mRNAs for surfactant proteins A, B, and C (SP-A, SP-B, and SP-C). Our results show that keratinocyte growth factor (KGF) induced a two- to threefold increase in steady-state levels of mRNAs for SP-A and SP-B, but had no effect on or decreased SP-C mRNA. The increase in SP-A mRNA was accompanied by an increase in SP-A protein. The effects of KGF were both dose and time dependent, and they could be neutralized by a monoclonal antibody against KGF. The effects of KGF were mimicked by acidic fibroblast growth factor, which will bind the KGF receptor. We conclude that KGF can support differentiation of alveolar type II cells as well as act as a mitogen, thus suggesting an important role for KGF in maintenance of the alveolar epithelium.

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Prevention of Paraquat Toxicity in Suspensions of Alveolar Type II Cells by Paraquat-Specific Antibodies

Human & Experimental Toxicology ◽

10.1177/096032719401300808 ◽

1994 ◽

Vol 13 (8) ◽

pp. 551-557 ◽

Cited By ~ 18

Author(s):

Nian Chen ◽

Mark R. Bowles ◽

Susan M. Pond

Keyword(s):

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Specific Antibodies ◽

Fab Fragments ◽

Paraquat Toxicity ◽

Specific Igg ◽

Energy Dependent

1 The herbicide, paraquat, is accumulated by the energy-dependent polyamine uptake pathway of alveolar type II cells. There it undergoes redox cycling that results in an amplified production of toxic reactive oxygen species and depletion of NADPH and other reducing equivalents. These processes account for the lung being the major target organ for paraquat toxicity. 2 We postulated that paraquat-specific antibodies would inhibit the uptake of the herbicide by type II cells and prevent its toxicity. Accordingly, we examined the effects of paraquat-specific monoclonal antibodies and Fab fragments on the uptake, efflux and cytotoxicity of 50 μM paraquat in suspensions of alveolar type II cells isolated from the rat. 3 The uptake of paraquat was linear over 40 min. Over this time, the uptake rate was inhibited significantly (% inhibition, 73-89) by IgG (25 or 50 μM) or Fab fragments (50 or 100 μM). 4 The apparent efflux rate of paraquat, studied over 16 h, was increased significantly from 0.12 h-1 for the control cells in medium to 0.17 h-1 by paraquat-specific Fab fragments but was unaffected by the specific IgG. 5 Cytotoxicity was determined by measuring the release of 51Cr from the cells. The cytotoxicity of 50 μM paraquat was decreased significantly (percent decrease, 56-80%) in the presence of specific antibodies. 6 These studies in vitro suggest some potential for immunotherapy in selected cases of paraquat poisoning.

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Distinct effects of SP-B and SP-C on the uptake of surfactant-like liposomes by alveolar cells in vivo and in vitro

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00054.2004 ◽

2004 ◽

Vol 287 (5) ◽

pp. L1056-L1065 ◽

Cited By ~ 10

Author(s):

D. L. H. Poelma ◽

L. J. Zimmermann ◽

W. A. van Cappellen ◽

J. J. Haitsma ◽

B. Lachmann ◽

...

Keyword(s):

Alveolar Macrophages ◽

Divalent Cations ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Cells ◽

Alveolar Type Ii Cells ◽

Lipid Ratio

The effects of surfactant protein B (SP-B) and SP-C on the uptake of surfactant-like liposomes by alveolar type II cells and alveolar macrophages were studied both in vivo and in vitro. In vivo, mechanically ventilated rats were intratracheally instilled with fluorescently labeled liposomes that had SP-B and/or SP-C incorporated in different concentrations. Consequently, the alveolar cells were isolated, and cell-associated fluorescence was determined using flow cytometry. The results show that the incorporation of SP-B does not influence the uptake, and it also does not in the presence of essential cofactors. The inclusion of SP-C in the liposomes enhanced the alveolar type II cells at a SP-C to lipid ratio of 2:100. If divalent cations (calcium and magnesium) were present at physiological concentrations in the liposome suspension, uptake of liposomes by alveolar macrophages was also enhanced. In vitro, the incorporation of SP-B affected uptake only at a protein-to-lipid ratio of 8:100, whereas the inclusion of SP-C in the liposomes leads to an increased uptake at a protein-to-lipid ratio of 1:100. From these results, it can be concluded that SP-B is unlikely to affect uptake of surfactant, whereas SP-C in combination with divalent cations and other solutes are capable of increasing the uptake.

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A bilayer tissue culture model of the bovine alveolus

F1000Research ◽

10.12688/f1000research.18696.1 ◽

2019 ◽

Vol 8 ◽

pp. 357

Author(s):

Diane Lee ◽

Mark Chambers

Keyword(s):

Epithelial Cells ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Host Pathogen Interactions ◽

Alveolar Type Ii Cells ◽

Permeable Membrane ◽

Host Pathogen

The epithelial lining of the lung is often the first point of interaction between the host and inhaled pathogens, allergens and medications. Epithelial cells are therefore the main focus of studies which aim to shed light on host-pathogen interactions, to dissect the mechanisms of local host immunity and study toxicology. If these studies are not to be conducted exclusively in vivo, it is imperative that in vitro models are developed with a high in vitro-in vivo correlation. We describe here a co-culture bilayer model of the bovine alveolus, designed to overcome some of the limitations encountered with mono-culture and live animal models. Our system includes bovine pulmonary arterial endothelial cells (BPAECs) seeded onto a permeable membrane in 24 well Transwell format. The BPAECs are overlaid with immortalised bovine alveolar type II epithelial cells and the bilayer cultured at air-liquid interface for 14 days before use; in our case to study host-mycobacterial interactions. Characterisation of novel cell lines and the bilayer model have provided compelling evidence that immortalised bovine alveolar type II cells are an authentic substitute for primary alveolar type II cells and their culture as a bilayer in conjunction with BPAECs provides a physiologically relevant in vitro model of the bovine alveolus. The bilayer model may be used to study dynamic intracellular and extracellular host-pathogen interactions, using proteomics, genomics, live cell imaging, in-cell ELISA and confocal microscopy. The model presented in this article enables other researchers to establish an in vitro model of the bovine alveolus that is easy to set up, malleable and serves as a comparable alternative to in vivo models, whilst allowing study of early host-pathogen interactions, currently not feasible in vivo. The model therefore achieves one of the 3Rs objectives in that it replaces the use of animals in research of bovine respiratory diseases.

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Mechanisms of Surfactant Exocytosis in Alveolar Type II Cells In Vitro and In Vivo

Physiology ◽

10.1152/physiologyonline.2001.16.5.239 ◽

2001 ◽

Vol 16 (5) ◽

pp. 239-243 ◽

Cited By ~ 5

Author(s):

Paul Dietl ◽

Thomas Haller ◽

Norbert Mair ◽

Manfred Frick

Keyword(s):

Surface Tension ◽

In Vitro Studies ◽

Alveolar Type ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Type Ii Cells ◽

Surfactant Secretion

Surfactant secretion must be regulated to maintain a low surface tension in the lung during various conditions such as exercise. In vitro studies reveal a slow, unique exocytotic process at the interface of stimulated and constitutive exocytosis. The exocytotic mechanisms and sites of regulation in vivo, however, are still poorly understood.

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