Mechanisms of suppression on cytokine expression in HTLV-1-infected T cell clones by aqueous humor

房水抑制HTLV-1感染T细胞克隆细胞因子表达的机制

• 批准号: 09671825
• 负责人: YOSHIMURA Koichi
• 金额: $ 1.98万
• 依托单位: Kurume 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-09671825/
• 关键词: HTLV-1; uveitis; aqueous humor; cytokine; suppressive factor; sFasL

In spite of constitutive overexpression of inflammatory cytokines in HTLV-1-infected T-cells, most cytokine genes are not expressed in the fresh infiltrating cells in the eye, suggesting a suppression of cytokine gene expression. To understand pathophysiology of HU, we characterized the suppressive factors in the aqueous humor (AH) and the mechanisms of suppression. AH dose-dependently suppressed the production of IFN-gamma and IL-6 by HTLV-l-infected T-cell clones and a cell line, Sez. Fractionation of AH by gel filtration column revealed four peaks of suppressive activity, which apparently corresponded molecular masses of a-MSH, VIP, MIF and TGF-beta. However, TGF-beta2, a major immunosuppressive factor in AH, did not show any suppressive activity. We demonstrated sFasL in AH by ELISA using NOKl and NOK3 anti-FasL mAbs and revealed neutralization of the suppression by NOK2 mAb. Crosslinking Fas selectively repressed the IFN-gamma mRNA expression. His-tagged sFasL showed dose-dependent suppression of IFN-gamma production and promoter activity in transient Luciferase assay. Suppression was evident at the concentration as low as 10 to l00pg/ml. Deletion and mutation analyses of IFN-gamma promoter identified a responsive element of 24 nucleotides to the Fas-mediated suppression, which contained binding motifs of YYl and an B-box. Mutation of the E-box in this element abrogated the suppression. Suppression was abolished by expression of dominant negative form of Daxx, an adaptor protein of Fas. The results indicated that sFasL is one of the suppressive factors of cytokine production in the eye, which would play a protective role against cytokine-mediated tissue damages. They also suggested a distinct function of sFasL from the membrane-bound from of FasL.

尽管炎性细胞因子在HTLV-1感染的T细胞中组成性过表达，但大多数细胞因子基因在眼睛中的新鲜浸润细胞中不表达，表明细胞因子基因表达受到抑制。为了了解HU的病理生理学，我们描述了房水（AH）中的抑制因子及其抑制机制。AH剂量依赖性地抑制HTLV-1感染的T细胞克隆和细胞系Sez.通过凝胶过滤柱对AH进行分级，显示出四个抑制活性峰，其显然对应于α-MSH、VIP、MIF和TGF-β的分子量。然而，TGF-β 2，一个主要的免疫抑制因子在AH，没有显示出任何抑制活性。我们使用NOK 1和NOK 3抗FasL mAb通过ELISA证明了AH中的sFasL，并揭示了NOK 2 mAb对抑制的中和作用。交联Fas选择性抑制IFN-γ mRNA的表达。在瞬时荧光素酶测定中，His标记的sFasL显示IFN-γ产生和启动子活性的剂量依赖性抑制。在低至10 - 100 pg/ml的浓度下抑制是明显的。IFN-γ启动子的缺失和突变分析鉴定了一个24个核苷酸的对Fas介导的抑制的响应元件，其包含YY 1和B-box的结合基序。该元件中E盒的突变消除了抑制。抑制被废除的显性负性形式的Daxx，Fas的衔接蛋白的表达。结果提示，sFasL是眼组织细胞因子产生的抑制因子之一，对眼组织损伤具有保护作用。他们还提出了一个不同的功能sFasL从膜结合形式的FasL。

项目成果

Ayako Ono: "Immunological and virological characterization of the primary infiltrating cells in the aqueous humor of human T-cell leukemia virus type-1 uveitis" Invest.Ophthalmol.Vis.Sci.38. 676-689 (1997)

Ayako Ono：“人 T 细胞白血病病毒 1 型葡萄膜炎眼房水中原代浸润细胞的免疫学和病毒学特征”Invest.Ophthalmol.Vis.Sci.38。

Mami Sakaguchi, Sunao Sugita, Kimitaka Sagawa, Kyogo Itoh, Manabu Mochizuki: "Cytokine production by T cells infiltrating in the eye of uveitiis patients." Japanese Journal of Ophthalmology. 42. 262-268 (1998)

Mami Sakaguchi、Sunao Sugita、Kimitaka Sakawa、Kyogo Itoh、Manabu Mochizuki：“浸润葡萄膜炎患者眼睛的 T 细胞产生细胞因子。”

Ayako Ono: "Provirus load in patients with human T-cell leukemia virus type I uveitis correlates with precedent Graves' disease and disease activities" Japanese Journal of Cancer Research. 89・6. 608-614 (1998)

小野绫子 (Ayako Ono)：“人类 T 细胞白血病病毒 I 型葡萄膜炎患者的原病毒载量与先前的格雷夫斯病和疾病活动相关”，日本癌症研究杂志 89・6 (1998)。

Yasuhiro Sato, Kinji Ito, Takashi Moritoyo, Yujiro Fujino, Kanjiro Masuda, Kazunari Yamaguchi, Manabu Mochizuki, Shuji Izumo, Mitsuhiro Osame, Toshiki Watanabe: "Human T-cell lymphotropic virus type 1 can infect primary rat ratinal glial cells and induce

Yasuhiro Sato、Kinji Ito、Takashi Moritoyo、Yujiro Fujino、Kanjiro Masuda、Kazunari Yamaguchi、Manabu Mochizuki、Shuji Izumo、Mitsuhiro Osame、Toshiki Watanabe：“人类 T 细胞嗜淋巴细胞病毒 1 型可以感染原代大鼠大鼠神经胶质细胞并诱导

望月 学: "局所T細胞と病態 : 眼" リウマチ科. 17・6. 588-592 (1997)

望月学：“局部T细胞和病理学：眼”风湿病学17・6（1997）。