Identification of molecules involved in inactivation of mast cells.

鉴定参与肥大细胞失活的分子。

• 批准号: 13557040
• 负责人: KITAMURA Toshio
• 金额: $ 8.83万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557040/
• 关键词: mast cell; NK receptor; allergy; paired Ig receptor; phosphatase; signal sequence trap; ITIMモチーフ; シグナルシークエンストラップ; 膜蛋白質; シグナル伝達

We searched for secreted and membrane proteins from cultured bone marrow-derived mouse mast cells using retrovirus-mediated signal sequence trap method SST-REX. From this screening, we identified a novel membrane protein harboring a homology with inhibitory NK receptors. This protein harbors a single immunoglobulin like domain in the extracellular domein and three or four ITIM inhibitory motifs in the intracellular domain, and was designated as LMIRl that stands for Jeukocyte-associated mono Ig receptor-l. From the data base search, we also identified LMIR-2-6 that resemble LMIR1, and isolated full-length cDNAs for these proteins. LMIRl and 3 harbors ITIM motifs in the intracellular domain, and upon ligation, they induced phosphorylation of phosphatases SHP1, SHP2, and SHIP, thereby inducing inhibitory signals. On the other hand, LMIR2, 4, 5, and 6 have short intracellular domain, and associate with activating receptors including DAP10, DAP12 and FcRg through the transmembrane domains. We have developed monoclonal antibodies against LMIR-1-6. To clarify the physiological roles of these molecules especially in the control of mast cell activation, we are now trying to identify the ligands for LMIR1 and 3, and to generate the knockout mice for LMIRI-3.

我们使用逆转录病毒介导的信号序列陷阱方法SST-Rex从培养的骨髓衍生的小鼠肥大细胞中搜索了分泌的和膜蛋白。通过此筛查，我们发现了一种具有抑制性NK受体同源性的新型膜蛋白。该蛋白具有在细胞外白蛋白中的单个免疫球蛋白类似的结构域和细胞内结构域中的三个或四个ITIM抑制基序，并被指定为代表二核细胞相关的单体IG受体LMIRL。从数据库搜索中，我们还确定了类似于LMIR1的LMIR-2-6，并为这些蛋白质分离了全长cDNA。 LMIRL和3个含有细胞内结构域中的ITIM基序，在结扎率后，它们诱导磷酸酶SHP1，SHP2和船舶的磷酸化，从而诱导抑制信号。另一方面，LMIR2、4、5和6具有短的细胞内结构域，并通过跨膜结构域（包括DAP10，DAP12和FCRG）与激活受体有关。我们已经开发了针对LMIR-1-6的单克隆抗体。为了阐明这些分子的生理作用，尤其是在控制肥大细胞激活中，我们现在正在尝试识别LMIR1和3的配体，并为LMIRI-3生成敲除小鼠。

项目成果

Nishiyama, C.: "Overproduction of PU.1 in mast cell progenitors : its effect on monocyte-and mast cell-specific gene expression."Biochem.Byophys.Res.Commun.. 313. 516-521 (2004)

Nishiyama, C.：“肥大细胞祖细胞中 PU.1 的过量产生：其对单核细胞和肥大细胞特异性基因表达的影响。”Biochem.Byophys.Res.Commun.. 313. 516-521 (2004)

Nakamura, T., Ouchida, R., Kodarna, T., Kawashima, T., Watanabe, S., Morimoto, C., Kitamura, T., Tanaka, H.: "Cytokine receptor common β subunit-mediated STAT5 activation confers NFκB activation in murinepro B cell line Ba/F3 cells."J. Biol. Chem.. 277. 6

Nakamura, T.、Ouchida, R.、Kodarna, T.、Kawashima, T.、Watanabe, S.、Morimoto, C.、Kitamura, T.、Tanaka, H.：“细胞因子受体常见 β 亚基介导的 STAT5 激活在 murinepro B 细胞系 Ba/F3 细胞中赋予 NFκB 激活作用。“J. Biol. Chem.. 277. 6

Ye, S.K., Agata, Y., Lee, H.C., Kurooka, H., Kitamura, T., Shimizu, A., Honjo, T., Ikuta, K.: "The IL-7 receptor controls the accessbility of the TCR γ locus by Stat5 and histone acetylation."Immunity. 15. 813-823 (2001)

Ye, S.K.、Agata, Y.、Lee, H.C.、Kurooka, H.、Kitamura, T.、Shimizu, A.、Honjo, T.、Ikuta, K.：“IL-7 受体控制 TCR 的可及性Stat5 和组蛋白乙酰化的 γ 位点。“Immunity. 15. 813-823 (2001)

Senga, T., Iwamoto, T., Kitamura, T., Miyake, Y., Hamaguchi, M.: "JAK/Stat3 dependent activation of RalGDS/Ral pathway in Ml mouse myeloid leukemic cells."J. Biol. Chem.. 276. 32678-32681 (2001)

Senga, T.、Iwamoto, T.、Kitamura, T.、Miyake, Y.、Hamaguchi, M.：“M1 小鼠骨髓白血病细胞中 RalGDS/Ral 途径的 JAK/Stat3 依赖性激活。”

Nagai, Y., Akashi, S., Nagafuku, M., Ogata, M., Iwakura, Y., Akira, S., Kitamura, T., Kosugi, A., Kimoto, M., Miyake, K.: "Essential role of MD-2 in LPS responsiveness and TLR4 distribution."Nat. Immunol.. 3. 667-672 (2002)

Nagai, Y.、Akashi, S.、Nagafuku, M.、Ogata, M.、Iwakura, Y.、Akira, S.、Kitamura, T.、Kosugi, A.、Kimoto, M.、Miyake, K.：