Targeting NLRP3 and staphylococcal pore-forming toxin receptors in human-induced pluripotent stem cell-derived macrophages

Seong H. Chow, Pankaj Deo, Amy T.Y. Yeung, Xenia P. Kostoulias, Yusun Jeon, Mei-Ling Gao, Azadeh Seidi, Françios Alwyn Benson Olivier, Sushmita Sridhar, Cara Nethercott, David Cameron, Avril A.B. Robertson, Remy Robert, Charles R. Mackay, Ana Traven, Zi-Bing Jin, Christine Hale, Gordon Dougan, Anton Y. Peleg, Thomas Naderer

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Staphylococcus aureus causes necrotizing pneumonia by secreting toxins such as leukocidins that target front-line immune cells. The mechanism by which leukocidins kill innate immune cells and trigger inflammation during S. aureus lung infection, however, remains unresolved. Here, we explored human-induced pluripotent stem cell-derived macrophages (hiPSC-dMs) to study the interaction of the leukocidins Panton–Valentine leukocidin (PVL) and LukAB with lung macrophages, which are the initial leukocidin targets during S. aureus lung invasion. hiPSC-dMs were susceptible to the leukocidins PVL and LukAB and both leukocidins triggered NLPR3 inflammasome activation resulting in IL-1β secretion. hiPSC-dM cell death after LukAB exposure, however, was only temporarily dependent of NLRP3, although NLRP3 triggered marked cell death after PVL treatment. CRISPR/Cas9-mediated deletion of the PVL receptor, C5aR1, protected hiPSC-dMs from PVL cytotoxicity, despite the expression of other leukocidin receptors, such as CD45. PVL-deficient S. aureus had reduced ability to induce lung IL-1β levels in human C5aR1 knock-in mice. Unexpectedly, inhibiting NLRP3 activity resulted in increased wild-type S. aureus lung burdens. Our findings suggest that NLRP3 induces macrophage death and IL-1β secretion after PVL exposure and controls S. aureus lung burdens.

Original languageEnglish
Number of pages15
JournalJournal of leukocyte biology
DOIs
Publication statusAccepted/In press - 12 Jun 2020

Keywords

  • inflammation
  • macrophage
  • NLRP3
  • pneumonia
  • staphylococcus
  • toxin

Cite this

Chow, S. H., Deo, P., Yeung, A. T. Y., Kostoulias, X. P., Jeon, Y., Gao, M-L., Seidi, A., Olivier, F. A. B., Sridhar, S., Nethercott, C., Cameron, D., Robertson, A. A. B., Robert, R., Mackay, C. R., Traven, A., Jin, Z-B., Hale, C., Dougan, G., Peleg, A. Y., & Naderer, T. (Accepted/In press). Targeting NLRP3 and staphylococcal pore-forming toxin receptors in human-induced pluripotent stem cell-derived macrophages. Journal of leukocyte biology. https://doi.org/10.1002/JLB.4MA0420-497R