Analysis of the function of membrane TNF-α

膜TNF-α的功能分析

• 批准号: 14570418
• 负责人: HORIUCHI Takahiko
• 金额: $ 1.92万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570418/
• 关键词: cytokine; anti-TNF-αtherapy; Crohn's disease; Rheumatoid arthritis; アポトーシス

TNF-□ antagonists are the. center of attention for their dramatic clinical efficacy in active chronic inflammatory diseases. In rheumatoid arthritis, both infliximab (chimeric anti-TNF-□ antibody) and etanercept(p75 TNF-□ receptor fusion protein) are highly effective while in Crohn's disease, only infliximab can induce clinical remission. As the differential clinical efficacy was likely to be caused by biological effects other than mere neutralization of soluble TNF-□, we here investigated reverse signaling through transmembrane TNF-□mTNF) induced by these drugs. Both infliximab and etanercept induced E-selectin expression on human Jurkat T cells stably transfected with mTNF, however only infliximab was able to induce IL-10 production, apoptosis, ROS accumulation and G1 cell cycle arrest. We next examined the element of the mTNF essential for these biological effects induced by infliximab. Cytoplasmic serine residues at positions 2,5 and 27 of mTNF were sequentially substituted to alanine by site-directed mutagenesis. Infliximab-induced apoptosis and cell cycle arrest were completely abrogated by substitution of all of these three cytoplasmic serine residues. In this study, we revealed that novel biological effects induced by infliximab and etanercept were mediated by reverse signaling of mTNF, which might explain the differential clinical efficacy of these anti-TNF-□ agents. We also clarified that cytoplasmic serine residues were critical for the signal transduction through mTNF.

TNF-□拮抗剂是。因其在活动性慢性炎症性疾病中的显著临床疗效而备受关注。在类风湿性关节炎中，英夫利西单抗（嵌合抗TNF-□抗体）和依那西普（p75 TNF-□受体融合蛋白）均高度有效，而在克罗恩病中，仅英夫利西单抗可诱导临床缓解。由于不同的临床疗效可能是由生物学效应引起的，而不仅仅是可溶性TNF-α的中和，因此我们在此研究了这些药物诱导的跨膜TNF-α的反向信号传导。英夫利西单抗和依那西普均能诱导mTNF稳定转染的人Jurkat T细胞表达E-选择素，但只有英夫利西单抗能诱导IL-10产生、细胞凋亡、ROS积累和G1期细胞阻滞。我们接下来检查了英夫利西单抗诱导的这些生物学效应所必需的mTNF元素。通过定点突变将mTNF的2、5和27位的胞质丝氨酸残基依次取代为丙氨酸。英夫利昔单抗诱导的细胞凋亡和细胞周期停滞完全废除了所有这三个细胞质丝氨酸残基的取代。在这项研究中，我们发现英夫利西单抗和依那西普诱导的新的生物学效应是由mTNF的反向信号传导介导的，这可能解释了这些抗TNF-□药物的不同临床疗效。我们还阐明了细胞质丝氨酸残基是通过mTNF的信号转导的关键。

项目成果

Harashima S, Tsukamoto H, et al.: "Osteoprotegerin and receptor activator of nuclear factor kBligand expression in fibroblast-like synoviocytes from RA and OA patients"Rheumatology. 43. 396-397 (2004)

Harashima S、Tsukamoto H 等人：“骨保护素和核因子 kBligand 受体激活剂在 RA 和 OA 患者的成纤维样滑膜细胞中的表达”风湿病学。

Harashima S, Tsukamoto H, et al.: "Osteoprotegerin and receptor activator of nuclear factor kBligand expression in fibroblast-like synoviocytes from RA and patients"Rheumatology. 43. 396-397 (2004)

Harashima S、Tsukamoto H 等人：“骨保护素和核因子 kBligand 受体激活剂在 RA 和患者的成纤维样滑膜细胞中的表达”风湿病学。

Koyama T, Tsukamoto H, et al.: "A novel polymorphism of the human APRIL gene is associated with systemic lupus erythematosus"Rheumatology. 42. 1-6 (2003)

Koyama T、Tsukamoto H 等人：“人类 APRIL 基因的一种新多态性与系统性红斑狼疮相关”风湿病学。

Mitoma H, Horiuchi T, et al.: "Binding activities of infliximab and etanercept to transmembrane tumor necrosis factor-α"Gastroenterology. 126(3). 934-935 (2004)

Mitoma H、Horiuchi T 等人：“英夫利昔单抗和依那西普与跨膜肿瘤坏死因子-α 的结合活性”Gastroenterology 126(3) 934-935 (2004)。

Mitoma H, Horiuchi T, Tsukamoto H.: "Binding activities of infliximab and etanercept to transmembrane tumor necrosis factor (TNF)-□."Gastroenterology. 126(3). 934-935 (2004)

Mitoma H、Horiuchi T、Tsukamoto H.：“英夫利昔单抗和依那西普与跨膜肿瘤坏死因子 (TNF)-□ 的结合活性。胃肠病学”126(3)。