Study of the effect of environmental chemicals on the high-affinity IgG receptor-mediated signal transudation of mest cells.

研究环境化学物质对 Mest 细胞高亲和力 IgG 受体介导的信号转出的影响。

• 批准号: 15590120
• 负责人: TESHIMA Reiko
• 金额: $ 2.18万
• 依托单位: National Intsitute of Health Sciences
• 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-15590120/
• 关键词: Canine Mast cells; High-affinity IgG receptor; Degranulation; cDNA; Canine IgG; Substance P; Environmental chemicals; ヒトIgG; カルシウム応答; タンパク質チロシンリン酸化

1.Canine cutaneous mastocytoma-derived (CM-MC) cells were activated both IgG- and IgE-mediated mechanisms. IgG-mediated protein tyrosine phosphorylation and Ca^<2+> influx were similar to those mediated by IgE. Protein kinase inhibitor staurosporine inhibited IgG-mediated [Ca^<2+>]i elevation and histamine release in a dose-dependent manner.2.The presence of high affinity IgG receptors (FcγRI) protein and mRNA in CM-MC cells was determined by immunoprecipitation and RT-PCR methods. Then, the cDNA sequence of FcγRIα subunit was identified. The entire canine FcγRIα cDNA was 1119bp long (GenBank accession number, AB 101519). The overall identity of the canine FcγRIa cDNA to its human and mouse counterparts was 84% and 78%, respectively.3.We transfected cDNA of FcγRIα to COS-7 cells to examine the binding of monomeric IgG to the cells. The cDNA of canine FcγRIα was cloned by RT-PCR and 5'/3'-RACE from total RNA of CM-MC cells. The cDNA was ligated to mammalian expression vector and transfected into COS -7 cells. The binding of IgG onto the COS -7 cells was detected by flow-cytometry.4.CM-MC was activated by Substance P (10-30 μM) and C5a (0.5-1μM) dose-dependently. [Ca^<2+>]i elevation and degranulation by these substances was inhibited by islet- activated protein (IAP). Therefore, the activation of these substances seemed to be dependent on G-protein-coupled mechanism5.In conclusion, CM-MC cells were useful for the study of allergic inflammation caused by IgG-dependent mechanisms. The environmental chemicals that affect Substance P and C5a production might influence allergic inflammation caused by IgG-dependent mast cell activation.

1.犬皮肤肥大细胞瘤来源的(CM-MC)细胞被IgG和IgE介导的机制激活，IgG介导的蛋白酪氨酸磷酸化和Ca 2+ 流入与IgE蛋白激酶抑制剂十字孢菌素介导的相似。以剂量依赖性方式抑制IgG介导的[Ca^2+]i升高和组胺释放。2.高亲和力IgG受体的存在通过免疫沉淀和RT-PCR方法测定了CM-MC细胞中的(FcγRI)蛋白和mRNA，然后鉴定了整个犬FcγRIα cDNA长1119bp(GenBank登录号，AB 101519)。犬FcγRIa cDNA与其人类和小鼠礼物的同一性分别为84%和78%。 3.我们将FcγRIα的cDNA转染至COS-7细胞以检查单体IgG与细胞的结合。通过RT-PCR克隆犬FcγRIα的cDNA并连接来自CM-MC细胞的总RNA的5'/3'-RACE。转染至哺乳动物表达载体并转染COS -7细胞，流式细胞术检测IgG与COS -7细胞的结合。4.CM-MC。 [Ca ^ 2+ ]i 被物质 P (10-30 μM) 和 C5a (0.5-1 μM) 激活，且这些物质的升高和脱粒被胰岛激活蛋白 (IAP) 抑制。这些物质的激活似乎依赖于G蛋白偶联机制。 总之，CM-MC细胞可用于研究由IgG依赖性机制引起的过敏性炎症。影响物质的环境化学物质。 P 和 C5a 的产生可能会影响 IgG 依赖性肥大细胞激活引起的过敏性炎症。

项目成果

Ryosuke Nakamura: "Presence and primary sequence of a high-affinity IgG receptor on canine mastocytoma (CM-MC) cells."Immunogenetics.. 55. 271-275 (2003)

Ryosuke Nakamura：“犬肥大细胞瘤 (CM-MC) 细胞上高亲和力 IgG 受体的存在和一级序列。”免疫遗传学.. 55. 271-275 (2003)

Canine Mast Cell Activation via Human IgG1 and IgG4

• DOI: 10.1159/000080659
• 发表时间: 2004-09
• 期刊: International Archives of Allergy and Immunology
• 影响因子: 2.8
• 作者: Yoshitaka Sato;R. Teshima;R. Nakamura;K. Takagi;Nobuo Sasaki;J. Sawada;S. Kitani

Presence and primary sequence of a high-affinity IgG receptor on canine mastocytoma (CM-MC) cells

犬肥大细胞瘤 (CM-MC) 细胞上高亲和力 IgG 受体的存在及其一级序列

• 发表时间: 2003
• 期刊: Immunogenetics 55
• 作者: R.Nakamura;Y.Sato;K.Takagi;N.Sasaki;J.Sawada;S.Kitani;R.Teshima
• 通讯作者: R.Teshima

Presence and primary sequence of a high-affinity IgG receptor on canine mastocytoma (CM-MC) cells.

犬肥大细胞瘤 (CM-MC) 细胞上高亲和力 IgG 受体的存在及其一级序列。

• 发表时间: 2003
• 期刊: Immunogenetics.. 55
• 作者: Y.Sato;R.Teshima;R.Nakamura;K.Takagi;N.Sasaki;J.Sawada;S.Kitani;Yoshitaka Sato;Ryosuki Nakamura et al.
• 通讯作者: Ryosuki Nakamura et al.

Calcium response and FcepsilonRI expression in bone marrow-derived mast cells co-cultured with SCG neurites.

与 SCG 神经突共培养的骨髓源性肥大细胞中的钙反应和 FcepsilonRI 表达。

• 发表时间: 2005
• 期刊: Biol.Pharm.Bull 28
• 作者: Ryosuke Nakamura;Akio Suzuki
• 通讯作者: Akio Suzuki