Abstract
The pro-inflammatory cytokine IL-1β is a secreted protein that is cleaved by caspase-1 during inflammasome activation upon recognition of internal and external insults to cells. Purinergic receptor P2X7 has been described to be involved in the release pathway of bioactive mature IL-1β by activated immune cells. Microparticle (MP) shedding has also been recently recognized as a manner of cytokine IL-1β release. However, the understanding of purinergic receptor roles in the MP-mediated IL-1β release process is still rudimentary. Gasdermin-D (GSDM-D), a protein involved in pyroptosis and inflammasome activation, has been recently described to be involved in the release of microparticles by virtue of its pore-forming ability. Hence, our current work is aimed to study the role of P2X7 in regulating GSDM-D-mediated microparticles and thereby bioactive IL-1β release. We provide evidence that cleaved functional IL-1β release in microparticles upon LPS stimulation is regulated by GSDM-D and P2X7 in a two-step fashion. GSDM-D activation first regulates release of IL-1β and P2X7 into microparticles. Then, microparticulate active P2X7 receptor then regulates the release of bioactive IL-1β encapsulated in microparticles to be able to target other cells inducing IL-8. Using an ATP model of stimulation, we further demonstrated that extracellular ATP stimulation to IL-1β containing LPS microparticles induces release of its content, which when subjected to epithelial cells induced IL-8. This effect was blocked by P2X7 inhibitor, KN62, as well as by IL-1RA. Taken together, our findings demonstrate for the first time the synergistic critical roles of GSDM-D and purinergic receptors in the regulation of microparticulate bioactive IL-1β release and induction of target cell responses.
References
Dinarello CA (1997) Interleukin-1. Cytokine Growth Factor Rev 8(4):253–265
Hugel B, Martinez MC, Kunzelmann C, Freyssinet JM (2005) Membrane microparticle: two sides of the coin. Physiology 20:22–27
Distler JH, Huber LC, Gay S, Distler O, Pesetsky DS (2006) Microparticles as mediators of cellular cross-talk in inflammatory disease. Autoimmunity 39(8):683–690
Mitra S, Exline M, Habyarimana F, Gavrilin M, Baker P, Masters SL, Wewers MD, Sarkar A (2018) Microparticulate caspase-1 regulates gasdermin-D and pulmonary vascular endothelial cell injury. Am J Respir Cell Mol Biol 59:56–64. https://doi.org/10.1165/rcmb.2017-0393OC
Qu Y, Franchi L, Nunez G, Dubyak GR (2007) Nonclassical IL-1 beta secretion stimulated by P2X7 receptors is dependent on inflammasome activation and correlated with exosome release in murine macrophages. J Immunol 179(3):1913–1925
MacKenzie A, Wilson HL, Kriss-Toth A, Dower SK, North A, Surprenant A (2001) Rapid secretion of interleukin-1 beta by microvesicle shedding. Immunity 8:825–835
Wewers MD, Sarkar A (2009) P2X7 receptor and macrophage function. Purinergic Signal 5:189–195
Lin HY, Lin PH, Wu SH, Yang LT (2015) Inducible expression of gasdermin A3 in the epidermis causes epidermal hyperplasia and skin inflammation. Exp Dermatol 24(11):897–899
Mitra S, Wewers MD, Sarkar A (2015) Mononuclear phagocyte-derived microparticulate caspase-1 induces pulmonary vascular endothelial cell injury. PLoS One 10(12):e0145607
Solle M, Labasi J, Perregaux DG, Stam E, Petrushova N, Koller BH, Griffiths RJ, Gabel CA (2001) Altered cytokine production in mice lacking P2X(7) receptors. J Biol Chem 276(1):125–132
Feng S, Fox D, Ming Man S (2018) Mechanisms of gasdermin family members in inflammasome signaling and cell death. J Mol Biol 430(18 Pt B):3068–3080
Schneider KS, Groß CJ, Dreier RF, Saller BS, Mishra R, Gorka O, Heilig R, Meunier E, Dick MS, Ćiković T, Sodenkamp J, Médard G, Naumann R, Ruland J, Kuster B, Broz P, Groß O (2017) The inflammasome drives GSDMD-independent secondary pyroptosis and IL-1 release in the absence of caspase-1 protease activity. Cell Rep 21(13):3846–3859
Sarkar A, Mitra S, Mehta S, Raices R, Wewers MD (2009) Monocyte derived microvesicles deliver a cell death message via encapsulated caspase-1. PLoS One 4(9):e7140
Saeki N, Kuwahara Y, Sasaki H, Satoh H, Shiroishi T (2007) Gasdermin (GSDM-D) localizing to mouse chromosome 11 is predominantly expressed in upper gastrointestinal tract but significantly suppressed in human gastric cancer cells. Mamm Genome 11(9):718–724
Russo HM, Rathkey J, Boyd-Tressler A, Katsnelson MA, Abbott DW, Dubyak GR (2016) Active caspase-1 induces plasma membrane pores that precede pyroptotic lysis and are blocked by lanthanides. J Immunol 197(4):1353–1367
McKechnie NM, King BCR, Fletcher E, Braun G (2006) Fas-ligand is stored in secretory lysosomes of ocular barrier epithelia and released with microvesicles. Exp Eye Res 83:304–314
Funding
This work was supported by NIH RO1 HL24325 to Sarkar and NIH RO1GM108928 to Sarkar/Wewers.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflicts of interest
Srabani Mitra declares that she has no conflict of interest.
Anasuya Sarkar declares that she has no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Mitra, S., Sarkar, A. Microparticulate P2X7 and GSDM-D mediated regulation of functional IL-1β release. Purinergic Signalling 15, 119–123 (2019). https://doi.org/10.1007/s11302-018-9640-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11302-018-9640-5