IMMUNE REGULATION BY THE P38 MAP KINASE PATHWAYS

P38 MAP 激酶途径的免疫调节

• 批准号: 15590434
• 负责人: TANAKA Nobuyuki
• 金额: $ 1.86万
• 依托单位: TOHOKU UNIVERSITY
• 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590434/
• 关键词: p38; mkk3; mkk6; macrophage; LPS; inflammatory-cytokines; 抗原提示細胞; T細胞

In order to analyze the functional significance of the p38 MAP kinase pathways, mkk3 and mkk6 knockout mice were subjected to analysis. To examine the role of the kinases in innate immunity, bone marrow derived macrophages (Mφ) were prepared from mkk3-/-mkk6+/-mice and control mice. When mkk3-/-mkk6+/-derived Mφ were stimulated by LPS, secretion of TNFα as well as IL-12 was severely impaired. Further, inflammatory chemical mediators were also examined. iNOS expression, as judged by the real time RT-PCR, was significantly reduced. These results suggest that p38 MAP kinas pathways play significant roles in innate immunity, polarization of T helper T cells and inflammations. To see if p38 MAP kinase pathways are also involved in cellular apoptosis, a fibloblastoid cell line with mkk3-/-mkk6-/-genotype was generated. This cell line manifested impaired p38 activation upon TNFα stimulation but not by UV exposure, suggesting the possible roles for mkk4 for the residual activation of p38. Apoptosis upon starvation was clearly inhibited in the cells, which corresponded well with the tumorigenic activity of the cell when injected subcutaneously to nude mice. Collectively these results indicated that p38 MAP kinases not only plays role in immune regulation but also in apoptosis and tumor growth. Finally I identified molecules that are phosphorylated by various cytokines with similar kinetics to the p38 MAP kinases, namely STAM1 and Hrs, two molecules involved in vesicular transport. STAM and Hrs forms a tight complex and the degradation of STAM1 was controlled but the presence of Hrs. Since Hrs is expressed in T cells and regulate T cell survival, I concluded that not only p38 but also Hrs and STAMs are involved in immune regulation including T cell survival/apoptosis.

为了分析p38 MAP激酶途径的功能意义，MKK3和MKK6敲除小鼠进行了分析。为了检查激酶在先天免疫中的作用，由MKK3 - / - MKK6 +/-小鼠和对照小鼠制备了骨髓衍生的巨噬细胞（Mφ）。当MKK3 - / - MKK6 +/-衍生的Mφ被LPS刺激时，TNFα的分泌以及IL-12的分泌严重受损。此外，还检查了炎症化学介质。根据实时RT-PCR判断的iNOS表达显着降低。这些结果表明，p38 MAP激酶途径在先天免疫学，T细胞的极化和炎症中起着重要作用。为了了解p38 MAP激酶途径是否也参与细胞凋亡，生成了带有MKK3 - / - MKK6 - / - 基因型的fibloblastoid细胞系。该细胞系在TNFα刺激后而非紫外线暴露表现出p38激活受损，这表明MKK4可能在p38的残留激活中起作用。饥饿时的凋亡清楚地抑制了细胞，当皮下注射对裸鼠时，这与细胞的致瘤活性很好。总的来说，这些结果表明，p38 MAP激酶不仅在免疫调节中起作用，而且在凋亡和肿瘤生长中也起着作用。最终，我鉴定了由各种细胞因子磷酸化的分子，其动力学与p38 MAP激酶相似，即Stam1和HRS，这是两个参与囊泡转运的分子。 Stam和HRS形成一个紧密的复合物，并且控制了Stam1的降解，但存在HRS。由于HRS在T细胞中表达并调节T细胞存活，因此我得出结论，不仅p38，还包括HRS和Stams，还参与了包括T细胞存活/凋亡在内的免疫调节。

项目成果

Encyclopedia of Endocrine Diseases

• DOI: 10.1016/c2016-1-01662-0
• 发表时间: 2004
• 作者: L. Martini

Hrs, a Mammalian Master Molecule in Vesicular Transport and Protein Sorting, Suppresses the Degradation of ESCRT Proteins Signal Transducing Adaptor Molecule land 2.

Hrs 是一种哺乳动物囊泡运输和蛋白质分选的主分子，可抑制 ESCRT 蛋白质信号转导接头分子的降解 2。

• 发表时间: 2005
• 期刊: The Journal of Biological Chemistry 280
• 作者: H.Kobayashi;N.Tanaka ほか
• 通讯作者: N.Tanaka ほか

Nobuyuki Tanaka(他9名): "Mechanism of p38 MAP kinase activation in vivo"Genes & Development. 17・16. 1969-1978 (2003)

Nobuyuki Tanaka（和其他 9 人）：“体内 p38 MAP 激酶激活机制”17・16 1969-1978（2003）。

Cytokines, constitutive secretion

细胞因子，组成性分泌

• 发表时间: 2004
• 期刊: Encyclopedia of Endocrine Diseases (Elsevier Inc.) 1
• 作者: Tanaka;N.;Sugamura K.
• 通讯作者: Sugamura K.

Hrs, a mammalian master molecule in vesicular transport and protein-sorting, suppresses the degradation of ESCRT proteins STAM1 and STAM2.

Hrs 是哺乳动物囊泡运输和蛋白质分选中的主分子，可抑制 ESCRT 蛋白 STAM1 和 STAM2 的降解。

• 发表时间: 2005
• 期刊: J.Biol.Chem. 280
• 作者: Kobayashi;H.;Tanaka;N.;^* Asao;H.;Miura;S.;Semura;K.;Ishii;N;Sugamura;K.
• 通讯作者: K.