Pulses of Class I PI3kinase activity identify the release and recapture of prey from neutrophil phagosomes

Clare Muir; Constantino Carlos  Reyes-Aldasoro; Felix E. Ellett; Tomasz K  Prajsnar; Yin X  Ho; Audrey  Bernut; Catherine A. Loynes; Stone Elworthy; Kieran A  Bowden; Ashley J  Cadbury; Lynne R. Prince; Jason S King; Alison M  Condliffe; Steven A  Renshaw

doi:10.1101/2024.02.29.582694

Pulses of Class I PI3kinase activity identify the release and recapture of prey from neutrophil phagosomes

Clare Muir, Constantino Carlos Reyes-Aldasoro, Felix E. Ellett, Tomasz K Prajsnar, Yin X Ho, Audrey Bernut, Catherine A. Loynes, Stone Elworthy, Kieran A Bowden, Ashley J Cadbury, Lynne R. Prince, Jason S King, Alison M Condliffe, Steven A Renshaw

Royal (Dick) School of Veterinary Studies

Research output: Working paper › Preprint

Abstract / Description of output

Class I PI3kinases coordinate the delivery of microbicidal effectors to the phagosome by forming the phosphoinositide lipid second messenger, phosphatidylinositol (3, 4, 5)-trisphosphate (PIP3). However, the dynamics of PIP3 in neutrophils during a bacterial infection are unknown. We have therefore developed an in vivo, live zebrafish infection model that enables visualisation of dynamic changes in Class 1 PI3kinases (PI3K) signalling on neutrophil phagosomes in real-time. We have identified that on approximately 12% of neutrophil phagosomes PHAkt-eGFP, a reporter for Class 1 PI3K signalling, re-recruits in pulsatile bursts. This phenomenon occurred on phagosomes containing
structurally and morphologically distinct prey, including Staphylococcus aureus and Mycobacterium abscessus, and was dependent on the activity of the Class 1 PI3K isoform, PI3kinase γ. Detailed imaging suggested that ‘pulsing phagosomes’ represent neutrophils transiently reopening and reclosing phagosomes. This finding challenges the concept that phagosomes remain closed after prey
engulfment and we propose that neutrophils occasionally use this alternative pathway of phagosome maturation to release phagosome contents and/or to restart phagosome maturation if digestion has stalled

Original language	English
Pages	1-33
Number of pages	33
DOIs	https://doi.org/10.1101/2024.02.29.582694
Publication status	Published - 29 Feb 2024

Publication series

Name	bioRxiv
Publisher	Cold Spring Harbor Laboratory Press

Keywords / Materials (for Non-textual outputs)

phagocytosis
neutrophils
zebrafish
phosphatidylinositol (3, 4, 5)-trisphosphate
phagosome maturation

Access to Document

10.1101/2024.02.29.582694

2024.02.29.582694v1.fullFinal published version, 1.2 MBLicence: Creative Commons: Attribution (CC-BY)

Cite this

Muir, C., Reyes-Aldasoro, C. C., Ellett, F. E., Prajsnar, T. K., Ho, Y. X., Bernut, A., Loynes, C. A., Elworthy, S., Bowden, K. A., Cadbury, A. J., Prince, L. R., King, J. S., Condliffe, A. M., & Renshaw, S. A. (2024). Pulses of Class I PI3kinase activity identify the release and recapture of prey from neutrophil phagosomes. (pp. 1-33). (bioRxiv). https://doi.org/10.1101/2024.02.29.582694

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title = "Pulses of Class I PI3kinase activity identify the release and recapture of prey from neutrophil phagosomes",

abstract = "Class I PI3kinases coordinate the delivery of microbicidal effectors to the phagosome by forming the phosphoinositide lipid second messenger, phosphatidylinositol (3, 4, 5)-trisphosphate (PIP3). However, the dynamics of PIP3 in neutrophils during a bacterial infection are unknown. We have therefore developed an in vivo, live zebrafish infection model that enables visualisation of dynamic changes in Class 1 PI3kinases (PI3K) signalling on neutrophil phagosomes in real-time. We have identified that on approximately 12% of neutrophil phagosomes PHAkt-eGFP, a reporter for Class 1 PI3K signalling, re-recruits in pulsatile bursts. This phenomenon occurred on phagosomes containingstructurally and morphologically distinct prey, including Staphylococcus aureus and Mycobacterium abscessus, and was dependent on the activity of the Class 1 PI3K isoform, PI3kinase γ. Detailed imaging suggested that {\textquoteleft}pulsing phagosomes{\textquoteright} represent neutrophils transiently reopening and reclosing phagosomes. This finding challenges the concept that phagosomes remain closed after preyengulfment and we propose that neutrophils occasionally use this alternative pathway of phagosome maturation to release phagosome contents and/or to restart phagosome maturation if digestion has stalled",

keywords = "phagocytosis, neutrophils, zebrafish, phosphatidylinositol (3, 4, 5)-trisphosphate, phagosome maturation",

author = "Clare Muir and Reyes-Aldasoro, {Constantino Carlos} and Ellett, {Felix E.} and Prajsnar, {Tomasz K} and Ho, {Yin X} and Audrey Bernut and Loynes, {Catherine A.} and Stone Elworthy and Bowden, {Kieran A} and Cadbury, {Ashley J} and Prince, {Lynne R.} and King, {Jason S} and Condliffe, {Alison M} and Renshaw, {Steven A}",

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AU - Reyes-Aldasoro, Constantino Carlos

AU - Ellett, Felix E.

AU - Prajsnar, Tomasz K

AU - Ho, Yin X

AU - Bernut, Audrey

AU - Loynes, Catherine A.

AU - Elworthy, Stone

AU - Bowden, Kieran A

AU - Cadbury, Ashley J

AU - Prince, Lynne R.

AU - King, Jason S

AU - Condliffe, Alison M

AU - Renshaw, Steven A

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AB - Class I PI3kinases coordinate the delivery of microbicidal effectors to the phagosome by forming the phosphoinositide lipid second messenger, phosphatidylinositol (3, 4, 5)-trisphosphate (PIP3). However, the dynamics of PIP3 in neutrophils during a bacterial infection are unknown. We have therefore developed an in vivo, live zebrafish infection model that enables visualisation of dynamic changes in Class 1 PI3kinases (PI3K) signalling on neutrophil phagosomes in real-time. We have identified that on approximately 12% of neutrophil phagosomes PHAkt-eGFP, a reporter for Class 1 PI3K signalling, re-recruits in pulsatile bursts. This phenomenon occurred on phagosomes containingstructurally and morphologically distinct prey, including Staphylococcus aureus and Mycobacterium abscessus, and was dependent on the activity of the Class 1 PI3K isoform, PI3kinase γ. Detailed imaging suggested that ‘pulsing phagosomes’ represent neutrophils transiently reopening and reclosing phagosomes. This finding challenges the concept that phagosomes remain closed after preyengulfment and we propose that neutrophils occasionally use this alternative pathway of phagosome maturation to release phagosome contents and/or to restart phagosome maturation if digestion has stalled

KW - phagocytosis

KW - neutrophils

KW - zebrafish

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KW - phagosome maturation

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Pulses of Class I PI3kinase activity identify the release and recapture of prey from neutrophil phagosomes

Abstract / Description of output

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Keywords / Materials (for Non-textual outputs)

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