Molecular analysis of anti-inflammatory effect of IL-10 and IL-4 on monocytes and neutrophils.

IL-10和IL-4对单核细胞和中性粒细胞抗炎作用的分子分析。

• 批准号: 09671120
• 负责人: OTSUKA Takeshi
• 金额: $ 1.66万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671120/
• 关键词: macrophage; neutrophil; interleukin-4; interleukin-10; cyclooxygenase; JAK-STAT; signal transduction; 単球

1) We investigated the effects of IL-4 and IL-1 on prostanoid synthesis in human neutrophils. Both cytokines inhibited their LPS-induced-PGE2 production, and their suppression was achieved by inhibiting cyclooxygense-2 mRNA expression. (Ref. 1)2) We previously reported that IL-4 and IL-l0 suppressed prostanoid synthesis in human monocytes.We questioned whether mitogen-activated protein kinase (MAPK) activation is involved in COX-2 expression in LPS-stimulated human monocytes. LPS induced the expression of COX-2 protein and COX-2 mRNA as well as the phospholylation and activation of extracellular signal-regulated protein kinase (ERK)2 and p38 MAPK in monocytes. The induction of COX-2 mRNA, COX-2 protein, and prostaglandin (PG) E2 by LPS was inhibited by the specific inhibitors of ERK2 and p38 MAPK.LPS-induced phosphorylation and activation of ERK2 was significantly inhibited by IL-4 and IL-10, while that of p38 MAPK was inhibited by IL-10, but not IL-4. These results suggested that the … More mechanisms of inhibition by IL-l0 and IL-4 of the LPS-induced expression of proinflammatory molecules could be ascribed to the regulatory effects of both cytokines on MAPK activation. (Ref. 2)3) We also studied whether some known STAT molecule is stimulated by LPS, based on the finding that a GAS motif sequence is conserved in the promoter regions of human, mouse, and rat COX-2 genes.Consequently, LPS induced activation of STAT5 in human monocytes, and this STATS activation occured in an indirect way via granulocyte-macrophage colony stimulating factor secreted by LPS- stimulated monocytes.Expression of COX-2 protein was partially reduced by treatment of anti-human GM-CSF Ab, Activation of STAT5 wa inhibited by either IL-10 or dexamethazone, but not by aspirin.IL-10 blocked activation of STATS indirectly by suppressing GM-CSF production, while Dex inhibited this activation both directly and indirectly.These results suggested that STAT5 plays an important role in activation of monocytes by LPS, and that STAT5 is another target for IL-10 and Dex to inhibit COX-2 expression. (Ref. 3)4) In rheumatoid arthritis patients, polymorphonuclear leukocytes are an important cellular source of prostanoids. To determine whether PMNs from RA patients exhibit different profiles in prostanoid synthesis compared to those from healthy donors, we studied 20 RA patients. Without stimulation, PGE2 production by PMNs was much higher in the RA patients than in the healthy controls. COX-2 protein was thought to be responsible for this increased PGE2 production. Neither IL-4 nor IL-10 affected the production. In conclusion, our results raise the possibility that PMNs from RA patients contribute, at least in part, to the pathogenesis of RA by producing COX-2-derived prostanoids in vivo. (Ref. 4)5) Since IL-10 was thought involved in pathogenesis of human diseases, we examined polymorphisms within human. IL-10 receptor cDNA gene sequenceby RT-PCR-RFLP.(Ref. 5) We further plan to examine the sensitivity of these polymorphisms for various immune disease. Less

1）我们研究了IL-4和IL-1对人神经粒细胞中前类动物合成的影响。两种细胞因子都抑制了其LPS诱导的PGE2产生，并且通过抑制环氧-2 mRNA表达来实现它们的抑制作用。 （参考文献1）2）我们先前报道说，IL-4和IL-L0抑制了人单核细胞中的前列腺素的合成。我们质疑有丝分裂原激活的蛋白激酶（MAPK）激活是否参与LPS刺激的人类单核细胞中的COX-2表达。 LPS诱导单核细胞中COX-2蛋白和Cox-2 mRNA的表达以及细胞外信号调节蛋白激酶（ERK）2和p38 mapk的磷酸化和激活。 LPS诱导Cox-2 mRNA，Cox-2蛋白和前列腺素E2受到ERK2和p38 MAPK.LPS诱导的抑制剂的抑制。LPS诱导的ERK2的磷酸化和ERK2的激活显着抑制了IL-4和IL-10，而P38 Map-10均受到P38 MAP的抑制。这些结果表明，在LPS诱导的促炎分子表达的IL-L0和IL-4抑制的更多机制可以分配给两种细胞因子对MAPK激活的调节作用。 （参考文献2）3）我们还研究了LP是否刺激了某些已知的Stat分子，这是基于以下发现，即气序序列是在人，小鼠和大鼠Cox-2基因的启动子区域中保守的，而LPS诱导的是，LPS诱导了人类单核细胞中的STAT5激活，以及这种统计数据通过granul刺激的gro刺激性，而granul刺激的分泌因素是Granul lly llanul lly ly ly ly ly ly ly ly ly ly ly ly ly ly ly ly ly ly，刺激的单核细胞。通过治疗抗人GM-CSF AB的处理，COX-2蛋白的表达部分降低，IL-10或dexamethazone抑制了STAT5 WA的激活，而AspiSin.IL.IL.IL-10并不能通过抑制GM-CSF的产生，同时抑制GM-CSF的产生，同时抑制了该统计数据的激活，同时又可以直接抑制这种激活。这些结果表明，STAT5在LPS激活单核细胞中起重要作用，而STAT5是IL-10和DEX抑制COX-2表达的另一个目标。 （参考3）4）在类风湿关节炎患者中，多形核白细胞是前列腺素的重要细胞来源。为了确定RA患者的PMN是否与健康捐助者相比是否暴露了前列腺素合成中的不同特征，我们研究了20名RA患者。没有刺激，RA患者的PMN产生PMN的生产要比健康对照组高得多。 COX-2蛋白被认为是PGE2产生增加的原因。 IL-4和IL-10都没有影响生产。总之，我们的结果提出了RA患者的PMN至少部分地通过在体内产生COX-2衍生的前列腺素来贡献RA的发病机理的可能性。 （参考文献4）5）由于认为IL-10参与了人类疾病的发病机理，因此我们检查了人类内部的多态性。 IL-10受体cDNA基因序列BRT-PCR-RFLP（参考文献5）我们进一步计划检查这些多态性对各种免疫疾病的敏感性。较少的

项目成果

大塚毅: "抗炎症作用を持つサイトカインによるRA制御の可能性" 医学のあゆみ. 182(9). 661-665 (1997)

Takeshi Otsuka：“通过具有抗炎作用的细胞因子控制 RA 的可能性”，《医学史》182(9)。

Niiro H: "Regulation by interleukin-10 and interleukin-4 of cyclooxygenase-2 expression in human neutrophils." Blood. 89(5). 1621-1628 (1997)

Niiro H：“白细胞介素 10 和白细胞介素 4 对人类中性粒细胞中环氧合酶 2 表达的调节。”

Niho Y: "Role of IL-10 in the crossregulation of prostaglandins and cytokines in monocytes." Acta Haematol. 99. 165-170 (1998)

Niho Y：“IL-10 在单核细胞中前列腺素和细胞因子交叉调节中的作用。”

Niiro H,Otsuka T,Izuhara K,Yamaoka K,Ohshima K,Tanabe T,Hara S,Nemoto Y,Tanaka Y,Nakashima H,Niho Y: "Regulation by interleukin-10 and interleukin-4 of cyclooxygenase-2 expression in human neutrophils." Blood. 89 (5). 1621-1628 (1997)

Niiro H、Otsuka T、Izuhara K、Yamaoka K、Ohshima K、Tanabe T、Hara S、Nemoto Y、Tanaka Y、Nakashima H、Niho Y：“白细​​胞介素 10 和白细胞介素 4 对人类环氧合酶 2 表达的调节

Niiro H,Otsuka T,Niho Y: "Down regulation by IL-10 of proinflammatory molecule expression." In : Niho Y (ed) : An Approach to Disease, -Immunology, Hematology, Oncology-Kyushu University Press 1-10. (1998)

Niiro H、Otsuka T、Niho Y：“IL-10 下调促炎分子表达。”