Mechanism of organophosphorus pesticides-induced inhibition of immune function by using perforin knockout mice.

有机磷农药诱导穿孔素基因敲除小鼠免疫功能抑制的机制。

• 批准号: 15590523
• 负责人: LI Qing
• 金额: $ 1.92万
• 依托单位: Nippon Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590523/
• 关键词: NK activity; CTL activity; LAK activity; Perforin; Granzyme; Granulysin; Perforin Knockout mice; Organophosphorus pesticide; perforin; 有機リン農薬; T細胞; マウス; DDVP; granzyme; granulysin; Fasligand; Fas pathway; FACS; NK; CTL; LAK; perforin

NK, LAK and CTL induce cell death in tumor or virus-infected target cells by two main mechanisms. The first mechanism is direct release of cytolytic granules that contain perforin, granzymes, and granulysin by exocytosis to kill target cells. The second mechanism is mediated by the Fas ligand (Fas-L)/Fas pathway.In the present syudy, we examined the effect that dimethyl 2,2-dichlorovinyl phosphate (DDVP), an organophosphorus pesticide has on NK, CTL and LAK activities of PKO mice in vitro using the Fas antigen-positive YAC-1 cell as a target. We found that DDVP significantly decreased NK, CTL and LAK activities in a dose-dependent manner, and that the CTL and LAK activities of PKO mice were significantly blocked by anti-FasL antibody, suggesting that DDVP and anti-FasL antibody have the same/similar mechanism of inhibiting LAK and CTL activities. We further found that DDVP decreases the expression of Fas antigen on YAC-1 cells, and the expression of FasL on LAK cells in a dose-dependent manner, respectively. Taken together, these findings indicate that the DDVP-induced inhibition of NK, LAK and CTL activities in PKO mice is mediated by the impairment of the FasL/Fas pathway. We also found that DDVP not only significantly inhibited activities of granzymes, but also decreased the intracellular level of perforin, granzyme A and granulysin, which was mediated by inducing degranulation of NK cells and by inhibiting the transcript of mRNA of perforin, granzyme A and granulysin. Taken together, these findings indicate that organophosphorus pesticides inhibited NK, LAK and CTL activities mediated at least by impairment of both the granule exocytosis and FasL/Fas pathways. We are grateful to Professor Krensky AM at Medical School of Stanford University for his helpful advice.

NK、LAK和CTL通过两种主要机制诱导肿瘤或病毒感染的靶细胞中的细胞死亡。第一种机制是通过胞吐作用直接释放含有穿孔素、颗粒酶和颗粒溶素的溶细胞颗粒以杀死靶细胞。本研究以Fas抗原阳性的YAC-1细胞为靶细胞，观察了有机磷农药2，2-二氯乙烯基磷酸酯（DDVP）对PKO小鼠NK、CTL和LAK活性的影响。结果表明，DDVP可剂量依赖性地降低PKO小鼠NK、CTL和LAK细胞活性，抗FasL抗体可显著阻断PKO小鼠的CTL和LAK细胞活性，提示DDVP和抗FasL抗体抑制LAK和CTL活性的机制相同或相似。DDVP可剂量依赖性地降低YAC-1细胞表面Fas抗原和LAK细胞表面FasL的表达。总之，这些发现表明DDVP诱导的PKO小鼠NK、LAK和CTL活性的抑制是通过FasL/Fas通路的损伤介导的。DDVP不仅能显著抑制NK细胞颗粒酶活性，而且还能降低NK细胞内穿孔素、颗粒酶A和颗粒溶素的水平，这种作用是通过诱导NK细胞脱颗粒和抑制穿孔素、颗粒酶A和颗粒溶素的mRNA转录而介导的。总之，这些研究结果表明，有机磷农药抑制NK，LAK细胞和CTL活性介导的颗粒胞吐和FasL/Fas途径的损害至少。我们感谢斯坦福大学医学院的Krensky AM教授提供的有益建议。

项目成果

Dimethyl 2,2-dichlorovinyl phosphate (DDVP) markedly inhibits activities of natural killer cells, cytotoxic T lymphocytes and lymphokine-activated killer cells via the Fas-ligand/Fas pathway in perform-knockout (PKO) mice.

二甲基 2,2-二氯乙烯基磷酸盐 (DDVP) 通过 Fas-配体/Fas 途径显着抑制执行基因敲除 (PKO) 小鼠中自然杀伤细胞、细胞毒性 T 淋巴细胞和淋巴因子激活的杀伤细胞的活性。

• 发表时间: 2004
• 期刊: Toxicology 204
• 作者: Li Q;Nakadai A;Takeda K. Kawada T.
• 通讯作者: Takeda K. Kawada T.

Dimethyl 2,2-dichlorovinyl phosphate (DDVP) markedly inhibits activities of natural killer cells, cytotoxic T lymphocytes and lymphokine-activated killer cells via the Fas-ligand/Fas pathway in perforin-knockout (PKO) mice

• DOI: 10.1016/j.tox.2004.05.019
• 发表时间: 2004-11-01
• 期刊: TOXICOLOGY
• 影响因子: 4.5
• 作者: Li, Q;Nakadai, A;Kawada, T
• 通讯作者: Kawada, T

Dimethyl 2,2-dichlorovinyl phosphate (DDVP) markedly decreases the expression of perforin, granzyme A and granulysin in human NK-92CI cell line

• DOI: 10.1016/j.tox.2005.05.018
• 发表时间: 2005-09-15
• 期刊: TOXICOLOGY
• 影响因子: 4.5
• 作者: Li, Q;Nakadai, A;Kawada, T
• 通讯作者: Kawada, T

The new mechanism of organophosphorus pesticides-induced inhibition of cytolytic activity of killer cells.

有机磷农药抑制杀伤细胞溶细胞活性的新机制

• 发表时间: 2006
• 期刊: Cellular & Molecular Immunology (in press)
• 作者: Li Q;Kawada T.
• 通讯作者: Kawada T.

Dimethyl 2, 2-dichlorovinyl phosphate (DDVP) markedly decreases the expression of perform, granzyme A and granulysin in human NK-92CI cell line.

磷酸二甲酯 (DDVP) 显着降低人 NK-92CI 细胞系中 perper、颗粒酶 A 和颗粒溶素的表达。

• 发表时间: 2005
• 期刊: Toxicology 213(1-2)
• 作者: Li Q;Nakadai A;Ishizaki M;Morimoto K;Ueda A;Krensky AM;Kawada T.
• 通讯作者: Kawada T.