Structure and function of the complement receptor type 2(CR2) in fish : regulation of the adaptive immunity by the complement system.

鱼类补体受体 2 型 (CR2) 的结构和功能：补体系统对适应性免疫的调节。

• 批准号: 13460092
• 负责人: YANO Tomoki
• 金额: $ 9.86万
• 依托单位: Kyushu University
• 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-13460092/
• 关键词: Fish; complement; innate immunity; C3d; adaptive immunity; CR2; receptor; diversity; コイ; 硬骨魚類; 分子クローニング

In mammals, an activation fragment(C3d) of the third complement component(C3) plays a crucial role in supporting nomal humoral immune responces, by stimulating B cells thrugh a specific receptor termed CR2. In the present study, we have examined if the adaptive immunity-regulatory effect is conserved in bony fish. First, we have analyzed physiolocial brakedown of carp C3 in serum activated by zymosan, and found that carp C3 generates a fragment equivalent to human C3d. It was also demonstrated that both C3-H1 and C3-S isoforms of carp C3 are fragmented into C3d. Next we have constructed expression vectors for fusion proteins of glutathione-S-transferase(GST) and C3d, and succeeded to prepare the recombinant proteins. Using the GST-C3d protein as a probe, we have attemted to detect receptors that recognize carp C3d fragment on carp peripheral lymphocytes. As a result, we observed significant and specific binding of GST-C3d to the lymphocytes. Anti-carp C3 rabbit IgG, but not EDTA, inhibited the binding, suggesting that carp lymphocytes carry receptors specific for C3d. The divalent-independent nature agrees well with that of mammalian CR2.

在哺乳动物中，第三补体成分（C3）的活化片段（C3 d）通过称为CR 2的特异性受体刺激B细胞，在支持正常体液免疫应答中起关键作用。在本研究中，我们已经研究了适应性免疫调节效应是否在硬骨鱼中是保守的。首先，我们分析了酵母多糖激活鲤鱼血清中C3的生理性制动，发现鲤鱼C3产生了一个与人C3 d相当的片段。鲤鱼C3的C3-H1和C3-S异构体均断裂成C3 d。其次，我们构建了谷胱甘肽-S-转移酶（GST）和C3 d融合蛋白的表达载体，并成功地制备了重组蛋白。以GST-C3 d蛋白为探针，在鲤鱼外周血淋巴细胞上检测识别鲤鱼C3 d片段的受体。结果，我们观察到GST-C3 d与淋巴细胞的显著和特异性结合。抗鲤鱼C3兔IgG，而不是EDTA，抑制结合，表明鲤鱼淋巴细胞携带C3 d的特异性受体。二价独立的性质与哺乳动物CR2的性质一致。

项目成果

矢野友紀, 中尾実樹: "硬骨魚類の補体の特性,「魚類の免疫系」,渡辺 翼編"恒星社厚生閣. 163 (2003)

Yuki Yano，Miki Nakao：“硬骨鱼补体的特征”，“鱼类免疫系统”，渡边翼编辑，Seiseisha Koukaku 163（2003）。

Kono T, Fujiki K, Nakao M, Yano T, Endo M, Sakai M.: "The immune responses of common carp, Cyprinus carpio L., injected with carp interleukin-1β gene"Journal of Interferon and Cytokine Research. 22. 413-419 (2002)

Kono T、Fujiki K、Nakao M、Yano T、Endo M、Sakai M.：“注射鲤鱼白细胞介素 1β 基因的鲤鱼的免疫反应”干扰素和细胞因子研究杂志 22. 413。 -419 (2002)

Nakao M, Hisamatsu S, Nakahara M, Kato Y, Smith SL, Yano T.: "Molecular cloning of the complement regulatory factor I isotypes from the common carp(Cyprinus carpio)"Immunogenetics. 54(in press). (2003)

Nakao M、Hisamatsu S、Nakahara M、Kato Y、Smith SL、Yano T.：“来自鲤鱼（Cyprinus carpio）的补体调节因子 I 同种型的分子克隆”免疫遗传学。

Nakao M, Matsumoto M, Nakazawa M, Fujiki K, Yano T: "DiversiLy of complement factor B/C2 in the common carp (Cyprinus carpio) : Three isotypes of B/C2-A expressed in different tissues"Dev. Comp. Immunol. 26(in press). (2002)

Nakao M、Matsumoto M、Nakazawa M、Fujiki K、Yano T：“鲤鱼（Cyprinus carpio）补体因子 B/C2 的多样性：在不同组织中表达的 B/C2-A 的三种同种型”Dev.

Nakao M, Fujiki K, Kondo M, Yano T: "Detection of complement receptors on head kidney phagocytes of the common carp (Cyprinus carpio)."Fisheries Science. 69. 927-933 (2003)

Nakao M、Fujiki K、Kondo M、Yano T：“鲤鱼（Cyprinus carpio）头肾吞噬细胞上补体受体的检测。”渔业科学。