Role of P2X7 receptor in Clostridium perfringens beta-toxin-mediated cellular injury

• Published in: Biochimica et biophysica acta Vol. 1850; no. 11; pp. 2159 - 2167
• Main Authors: Nagahama, Masahiro, Seike, Soshi, Shirai, Hidenori, Takagishi, Teruhisa, Kobayashi, Keiko, Takehara, Masaya, Sakurai, Jun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2015
• Subjects: ADAM Proteins - physiology; ADAM10 Protein; adenosine diphosphate; adenosine monophosphate; adenosine triphosphate; Amyloid Precursor Protein Secretases - physiology; Animals; antagonists; Bacterial Toxins - toxicity; C. perfringens beta-toxin; Calcium - metabolism; cell death; Clostridium perfringens; cytotoxicity; enterotoxemia; HEK293 Cells; Humans; lipids; Membrane Proteins - physiology; Mice; Mice, Inbred ICR; necrotic enteritis; Oligomer formation; P2X7 receptor; Pore-forming toxin; receptors; Receptors, Purinergic P2X7 - physiology; RNA, Small Interfering - pharmacology; Rosaniline Dyes - pharmacology; small interfering RNA; C. perfringens beta-toxin; Oligomer formation; Pore-forming toxin; P2X7 receptor; P2X receptor
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2015.08.011

Clostridium perfringens beta-toxin is a pore-forming toxin (PFT) and an important agent of necrotic enteritis and enterotoxemia. We recently reported that beta-toxin strongly induced cell death in THP-1 cells via the formation of oligomers. We here describe that the P2X7 receptor, which is an ATP receptor, interacts with beta-toxin. We tested the role of P2X7 receptor in beta-toxin-induced toxicity using specific inhibitors, knockdown of receptor, expression of the receptor and interaction by dot-blot assay. The potency of P2X7 receptor was further determined using an in vivo mouse model. Selective P2X7 receptor antagonists (oxidized ATP (o-ATP), oxidized ADP, and Brilliant Blue G (BBG)) inhibited beta-toxin-induced cytotoxicity in THP-1 cells. o-ATP also blocked the binding of beta-toxin to cells. The P2X7 receptor and beta-toxin oligomer were localized in the lipid rafts of THP-1 cells. siRNA for the P2X7 receptor inhibited toxin-induced cytotoxicity and binding of the toxin. In contrast, the siRNA knockdown of P2Y2 or P2Y6 had no effect on beta-toxin-induced cytotoxicity. The addition of beta-toxin to P2X7-transfected HEK-293 cells resulted in binding of beta-toxin oligomer. Moreover, beta-toxin specifically bound to immobilized P2X7 receptors in vitro and colocalized with the P2X7 receptor on the THP-1 cell surface. Furthermore, beta-toxin-induced lethality in mice was blocked by the preadministration of BBG. The results of this study indicate that the P2X7 receptor plays a role in beta-toxin-mediated cellular injury. P2X7 receptor is a potential target for the treatment of C. perfringens type C infection. •We demonstrate that P2X7 receptor is a target molecule of beta-toxin.•Beta-toxin-induced lethality in mouse is blocked by P2X7 receptor antagonist.•P2X7 receptor is a new therapeutic target for the treatment of C. perfringens type C infection.