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

• DOI: 10.1002/jlb.4ma0420-497r
• Publication Date: 2020-06-12T23:48:57Z
• Authors: Seong H Chow; Pankaj Deo; Amy T Y Yeung; Xenia P Kostoulias; Yusun Jeon; Mei-Ling Gao; Azadeh Seidi; Françios Alwyn Be...; 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
• Keywords: 0301 basic medicine; Bacterial Toxins; Induced Pluripotent Stem Cells; Interleukin-1beta; Exotoxins; macrophage; Monocytes; 1307 Cell Biology; Mice; 03 medical and health sciences; NLRP3; Bacterial Proteins; Leukocidins; NLR Family, Pyrin Domain-Containing 3 Protein; pneumonia; Animals; Humans; Gene Knock-In Techniques; toxin; Lung; Cells, Cultured; 2403 Immunology; CD11b Antigen; Macrophages; Cell Differentiation; 3. Good health; Mice, Inbred C57BL; inflammation; 2723 Immunology and Allergy; Leukocyte Common Antigens; CRISPR-Cas Systems; staphylococcus

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