| Project/Area Number |
17590406
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Tokushima Bunri University |
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Principal Investigator |
NAGAHAMA Masahiro Tokushima Bunri University, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助教授 (40164462)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Keiko Tokushima Bunri University, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (90170315)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | 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 |
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| Research Abstract |
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.
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