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Inflammatory Response and Barrier Properties of a New Alveolar Type 1-Like Cell Line (TT1)

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Abstract

Purpose

To evaluate the inflammatory response and barrier formation of a new alveolar type 1-like (transformed type I; TT1) cell line to establish its suitability for toxicity and drug transport studies.

Methods

TT1 and A549 cells were challenged with lipopolysaccharide (LPS). Secretion of inflammatory mediators was quantified by ELISA. The barrier properties of TT1 cells were evaluated by transepithelial electrical resistance (TEER), fluorescein sodium (flu-Na) apparent permeability (P app) and staining of zona occludens-1 (ZO-1).

Results

LPS stimulated similar levels of secretion of IL-6 and IL-8 in TT1 and A549 cells. TNF-α was not produced by either cell line. In contrast to A549 cells, TT1 cells did not secrete SLPI or elafin. TT1 cells produced maximal TEER of ~55 Ω cm2 and flu-Na P app of ~6.0 × 10−6 cm/s. ZO-1 staining was weak and discontinuous. Attempts to optimise culture conditions did not increase the barrier properties of the TT1 cell layers.

Conclusions

The TT1 cell line models the alveolar inflammatory response to LPS challenge and provides a valuable complement to cell lines currently used in toxicity assays. However, under the experimental conditions used the TT1 cell line did not form the highly restrictive tight junctions which exist in vivo.

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Abbreviations

AT1:

alveolar type 1

AT2:

alveolar type 2

flu-Na:

fluorescein sodium

P app :

apparent permeability coefficient

SLPI:

secretory leukoprotease inhibitor

TEER:

transepithelial resistance

TT1:

transformed type 1

ZO-1:

zona occludens protein-1

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Acknowledgements

EHJ van den Bogaard was supported by a grant from the Astma Fonds, The Netherlands. AJ Thorley was funded by Research Councils UK.

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Authors and Affiliations

  1. Pharmaceutical Science Division, King’s College London, Franklin-Wilkins Building, London, UK

    Ellen H. J. van den Bogaard, Lea Ann Dailey & Ben Forbes

  2. Radboud University Nijmegen, Nijmegen, The Netherlands

    Ellen H. J. van den Bogaard

  3. Lung Cell Biology, Section of Airways Disease, National Heart and Lung Institute, Imperial College, London, UK

    Andrew J. Thorley & Teresa D. Tetley

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  1. Ellen H. J. van den Bogaard
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Correspondence to Ben Forbes.

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van den Bogaard, E.H.J., Dailey, L.A., Thorley, A.J. et al. Inflammatory Response and Barrier Properties of a New Alveolar Type 1-Like Cell Line (TT1). Pharm Res 26, 1172–1180 (2009). https://doi.org/10.1007/s11095-009-9838-x

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  • DOI: https://doi.org/10.1007/s11095-009-9838-x

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