Structural Analysis and Mechanisms of Action of Various Inflammatory Cytokines

多种炎症细胞因子的结构分析及作用机制

• 批准号: 63440085
• 负责人: OSAWA Toshiaki
• 金额: $ 14.14万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63440085/
• 关键词: lymphokine; macrophage activating factor; macrophage migration inhibitory factor; lymphotoxin; tumor necrosis factor; interleukin-2; natural killer cell activating factor; マクロファ-ジ走化性因子; ナチュラルキラ-細胞活性化因子; マクロファージ走化性因子; ナチュラルキラー細胞活性化因子; マクロファージ遊走阻

1. Purification and structural analysis of inflammatory cytokines(a) Macrophage chemotactic factor : We purified a low molecular weight peptide having strong macrophage chemotactic activity and elucidated its amino acid sequence. Furthermore, we synthesized this peptide and observed that the synthetic peptide also has strong macrophage chemotactic activity.(b) Macrophage migration inhibitory factor (MIF) : We purified two kinds of MIF form the culture supernatant of human T cell hybridomas. We elucidated partial amino acid sequences of these two factors and found that these factors are novel lymphokines.(c) Macrophage activation factor for tumoricidal activity (MAF-C) : We purified MAF-C from the culture supernatant of human T cell hybridomas. Macrophage activating activity of the purified factor was completely inhibited by the addition of anti-IL-2 antibody, suggesting that MAF-C is actually IL-2 or an antigenically related substance with IL-2. Then, we analyzed macrophage activating of recombinant IL-2.2. Mechanism of action of tumor necrosis factorWe found that activated macrophages kill tumor cells in vivo which are resistant to tumor necrosis factor in vitro. Therefore, we analyzed the mechanism of tumoricidal activity of activated macrophages, and we found that NO produced by activated macrophages depending on the presence of arginine syn ergistically acts on tumor cells with tumor necrosis factor.

1.炎性细胞因子的纯化和结构分析（a）巨噬细胞趋化因子：我们纯化了具有强巨噬细胞趋化活性的低分子量肽，并阐明了其氨基酸序列。此外，我们合成了这种肽，并观察到合成的肽也具有很强的巨噬细胞趋化活性。(b)巨噬细胞移动抑制因子（MIF）：我们从人T细胞杂交瘤细胞培养上清中纯化了两种MIF。我们对这两个因子的部分氨基酸序列进行了分析，发现它们是新的淋巴因子。(c)巨噬细胞杀伤活性活化因子（MAF-C）：我们从人T细胞杂交瘤的培养上清液中纯化了MAF-C。纯化的因子的巨噬细胞活化活性被完全抑制通过添加抗IL-2抗体，表明MAF-C实际上是IL-2或与IL-2抗原相关的物质。然后，我们分析了重组IL-2.2的巨噬细胞活化。肿瘤坏死因子的作用机制我们发现活化的巨噬细胞在体内杀死肿瘤细胞，而在体外对肿瘤坏死因子具有抗性。因此，我们分析了活化的巨噬细胞的杀肿瘤活性的机制，并且我们发现活化的巨噬细胞依赖于精氨酸的存在而产生的NO与肿瘤坏死因子协同作用于肿瘤细胞。

项目成果

Naiki,M.: "Immunomodulatory effects of neurotropin through the recovery of interleukin 2 production in autoimmune(NZB x NZW)F_1 mice." Int.J.Immunopharmac.11. 663-671 (1989)

Naiki,M.：“神经营养素通过恢复自身免疫 (NZB x NZW)F_1 小鼠中白细胞介素 2 的产生而发挥免疫调节作用。”

Higuchi, M: "Tumoricidal activity of lymphotoxin (tumornecrosis factor bata) in vivo : its effects on macrophages." J. Biol. Response Mod.7. 619-630 (1988)

Higuchi, M：“体内淋巴毒素（肿瘤坏死因子 bata）的杀肿瘤活性：其对巨噬细胞的影响。”

Miyazaki,H.: "Enhancement of antigenーpresenting function of dendritic cells with culture supernatants of mouse peritoneal macrophage stimulated with certain particulate substances." Microbiol.Immunol.32. 1033-1042 (1988)

Miyazaki, H.：“用某些颗粒物质刺激的小鼠腹膜巨噬细胞的培养物上清液增强树突状细胞的抗原呈递功能。” Microbiol.Immunol.32 (1988)。

M.Higuchi: "TNFーmediated cytotoxicity.Importance of intracellular cGMP level for determining TNFーsensitivity" Molecular Immunology.

M. Higuchi：“TNF 介导的细胞毒性。细胞内 cGMP 水平对于确定 TNF 敏感性的重要性”分子免疫学。

Miyazaki, H.: "Enhancement of antigen-presenting function of dendritic cells with culture supernatants of mouse peritoneal macrophage stimulated with certain particulate substances." Micobiol. Immunol.32. 1033-1042 (1988)

Miyazaki, H.：“用某些颗粒物质刺激的小鼠腹膜巨噬细胞的培养上清液增强树突状细胞的抗原呈递功能。”