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Timothy Grass Pollen Induces Spatial Reorganisation of F-Actin and Loss of Junctional Integrity in Respiratory Cells

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Abstract

Grass pollens have been identified as mediators of respiratory distress, capable of exacerbating respiratory diseases including epidemic thunderstorm asthma (ETSA). It is hypothesised that during thunderstorms, grass pollen grains swell to absorb atmospheric water, rupture, and release internal protein content to the atmosphere. The inhalation of atmospheric grass pollen proteins results in deadly ETSA events. We sought to identify the underlying cellular mechanisms that may contribute towards the severity of ETSA in temperate climates using Timothy grass (Phleum pratense). Respiratory cells exposed to Timothy grass pollen protein extract (PPE) caused cells to undergo hypoxia ultimately triggering the subcellular re-organisation of F-actin from the peri junctional belt to cytoplasmic fibre assembly traversing the cell body. This change in actin configuration coincided with the spatial reorganisation of microtubules and importantly, decreased cell compressibility specifically at the cell centre. Further to this, we find that the pollen-induced reorganisation of the actin cytoskeleton prompting secretion of the pro-inflammatory cytokine, interleukin-8. In addition, the loss of peri-junctional actin following exposure to pollen proteins was accompanied by the release of epithelial transmembrane protein, E-cadherin from cell–cell junctions resulting in a decrease in epithelial barrier integrity. We demonstrate that Timothy grass pollen regulates F-actin dynamics and E-cadherin localisation in respiratory cells to mediate cell–cell junctional integrity highlighting a possible molecular pathway underpinning ETSA events.

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Acknowledgements

The authors would like to thank Maree Svolos (Respiratory Technology, Woolcock Institute of Medical Research, Glebe, Australia) for her helpful discussions.

Funding

Funding issued by Australian Research Council; ARC DP190100376, ARC DP190101973. Australian Research Council,DP190100376

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Authors and Affiliations

  1. Respiratory Technology, Woolcock Institute of Medical Research, Sydney, Australia

    Peta Bradbury, Aylin Cidem, Hui Xin Ong & Daniela Traini

  2. Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Sydney, Australia

    Hui Xin Ong & Daniela Traini

  3. School of Mathematical and Physical Sciences, University of Technology Sydney, Sydney, Australia

    Hadi Mahmodi & Irina Kabakova

  4. School of Biomedical Science, Centre for Immunology and Infection Control, Queensland University of Technology, Brisbane, Australia

    Janet M. Davies

  5. School of Chemical Engineering, UNSW, Sydney, Australia

    Patrick T. Spicer & Stuart W. Prescott

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Contributions

PB and AC contributed equally towards concept development, data curation, manuscript preparation, and figure preparation. HM and IK conducted Brillouin microscopy data curation and analysis. HO, DT, JMD, IK, PS, and SP provided critical feedback.

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Correspondence to Hui Xin Ong or Daniela Traini.

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Bradbury, P., Cidem, A., Mahmodi, H. et al. Timothy Grass Pollen Induces Spatial Reorganisation of F-Actin and Loss of Junctional Integrity in Respiratory Cells. Inflammation 45, 1209–1223 (2022). https://doi.org/10.1007/s10753-021-01614-9

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