Activation of calcineurin signal transduction by Clostridium perfingens beta-toxin in HL-60 cell

产气荚膜梭菌 β-毒素在 HL-60 细胞中激活钙调神经磷酸酶信号转导

• 批准号: 17590406
• 负责人: NAGAHAMA Masahiro
• 金额: $ 2.3万
• 依托单位: Tokushima Bunri University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590406/
• 关键词: Clostridium perfrigens; Beta-toxin; HL-60 cells; Phospholipase C; Calcineurin; Cyclosporin A; ラフト; 血球系細胞; ホスフォリパーゼC; オリゴマー形成

Clostridium perfringens beta-toxin is an important agent of necrotic enteritis. Of the 10 cell lines tested, only HL-60 cell line was susceptible to beta-toxin. The toxin induced swelling and lysis of the cells. The toxin induced production of diacylglycerol and inositol-3-phosphate (IP3) in a dose-and a time-dependent manner through the activation of phospholipase C (PLC) when incubated with HL-60 cells at 37 ℃, the toxin induced cell swelling were suppressed by Ca^<2+> antagonists (TMB-8,BAPTA-AM) and calcineurin (CN) inhibitor (cyclosporin A) but not by Ca^<2+> entry blocker (verapamil) and PKC inhibitor. When Fura-2-AM-treated HL-60 cells were incubated with the toxin in the absence of extracelluar Ca^<2+>, the cells increased intracellular Ca^<2+> in a dose-and a time-dependent manner. The results suggested that the toxin induced lysis of cells is closely related the intracelluar Ca^<2+> mobilization. Treatment of HL-60 cells with beta-toxin resulted in the activation of CN activity within 5 min. Cyclosporin A inhibited the toxin-induced increase of CN activity. An important intracelluar substrate for CN is NFTA (nuclear factor of activated T-cells). Activation of CN induced the translocation of NFTA to nucleus. The toxin stimulated the nuclear translocation of NFAT in HL-60 cells. The data indicated that beta-toxin-induced cell swelling is dependent on the Ca^<2+>/CaM-dependent CN through activity of PLC.

产气荚膜梭菌β毒素是坏死性肠炎的重要病原体。在所测试的10个细胞系中，仅HL-60细胞系对β-毒素敏感。毒素引起细胞肿胀和溶解。在37 ℃下，毒素通过激活磷脂酶C（PLC）诱导HL-60细胞产生甘油二酯和肌醇-3-磷酸（IP 3），并呈剂量和时间依赖性，Ca^<2+>拮抗剂可抑制毒素诱导的细胞肿胀（TMB-8，BAPTA-AM）和钙调磷酸酶（CN）抑制剂（环孢菌素A），但不被Ca^2+进入阻断剂（维拉帕米）和PKC抑制剂所抑制。当Fura-2-AM处理过的HL-60细胞在细胞外Ca^<2+>缺乏的情况下与毒素一起孵育时，细胞内Ca^<2+>以剂量和时间依赖的方式增加。结果提示，毒素诱导的细胞溶解与细胞内Ca^2+动员密切相关。用β-毒素处理HL-60细胞后，CN活性在5 min内被激活，环孢菌素A可抑制毒素诱导的CN活性的增加。CN的一种重要的细胞内底物是NFTA（活化T细胞的核因子）。激活CN可诱导NFTA向细胞核转位。该毒素刺激HL-60细胞NFAT核转位。这些数据表明，β-毒素诱导的细胞肿胀依赖于通过PLC活性的Ca^<2+>/CaM依赖性CN。

项目成果

Clostridium perfringens beta-toxin; characterization and action

产气荚膜梭菌β-毒素；

• 发表时间: 2006
• 期刊: J.Toxicol.Toxin Rev. 25
• 作者: J.Sakurai;M.Nagahama
• 通讯作者: M.Nagahama

The signal transduction mechanism involved in Clostridium perfringens alpha-toxin-induced superoxide anion generation in rabbit neutrophils

产气荚膜梭菌α毒素诱导兔中性粒细胞超氧阴离子产生的信号转导机制

• 发表时间: 2006
• 期刊: Infect. Immun. (In press)
• 作者: M.Oda;S.Ikari;T.Matsuno;Y.Morimune;M.Nagahama;J.Sakurai
• 通讯作者: J.Sakurai

Signal transduction mechanism involved in Clostridium perfringens alpha-toxin-induced superoxide anion generation in rabbit neutrophils

• DOI: 10.1128/iai.74.5.2876-2886.2006
• 发表时间: 2006-05-01
• 期刊: INFECTION AND IMMUNITY
• 影响因子: 3.1
• 作者: Oda, M;Ikari, S;Sakurai, J
• 通讯作者: Sakurai, J

Oligomerization of Clostridium perfringens epsilon-toxin is dependent upon membrane fluidity in liposomes.

产气荚膜梭菌ε-毒素的寡聚化取决于脂质体中的膜流动性。

• 发表时间: 2006
• 期刊: Biochemistry 45・1
• 作者: Tumurkhuu G;Koide N;Takahashi K;Hassan F;Islam S;Ito H;Mori I;Yoshida T;Yokochi T.;Masahiro Naganuma
• 通讯作者: Masahiro Naganuma

Role of tyrosine-57 and -65 in membrane-damaging and sphingomyelinase activities of Clostridium perfringens alpha-toxin

酪氨酸57和-65在产气荚膜梭菌α毒素膜损伤和鞘磷脂酶活性中的作用

• 发表时间: 2005
• 期刊: Biochim. Biophys. Acta 1762
• 作者: M.Nagahama;A.Otsuka;J.Sakurai.
• 通讯作者: J.Sakurai.