In vitroexpansion or induction of regulatory T cells from olcerative colitis patients

溃疡性结肠炎患者调节性 T 细胞的体外扩增或诱导

• 批准号: 18590685
• 负责人: NAKAMURA Kazuhiko
• 金额: $ 2.49万
• 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: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590685/
• 关键词: inflammatory bowel disease; regulatory T cell; immunology

CD4^+CD25^+ regulatory T cells (Treg) possess broad range of immunoregulatory activity which regulates autoimmune disorders including inflammatory bowel diseases in animal models. Aiming the clinical application of Treg for the treatment of ulcerative colitis (UC), we investigated whether Treg can be expanded or induced in vitro. We stimulated Tregs isolated from the leukapheresis products from UC patients with anti-CD3/anti-CD28-bearing beads and IL-2. Treg could be increased 18 times in 10 days culture. Cultured Treg maintained the expression of a Treg-specific marker, FOXP3 and significantly suppressed the CD4^+ T cell-proliferation upon coculture. Thus, Treg can be expanded in vitro.Next, we tested whether Treg can be induced in vitro CD4^+CD255non-Treg were stimulated and cultured in the presence of TGF-β1. Cultured cells expressed FOXP3 and suppressed the CD4^+ T cell-proliferation upon coculture. Therefore, Treg can be even induced from non-Treg in vitro.Then, we investigate whether induced Treg are able to suppress intestinal inflammation. We induced Treg from CD4^+ T cells stimulating with anti-CD3/anti-CD28 and IL-2 in the presence of rapamycin (RAPA). When CD4^+ T cells were stimulated in the presence of RAPA, 17% was CD25^+FoxP3^+ at the end of 21-day culture. In contrast, few cells stimulated without RAPA expressed CD25 and FoxP3. CB-17 Scid mice were transferred with CD4^+CD62L^+CD25^-T cells, which developed chronic persistent colitis. Co-transfer of CD4^+ T cells stimulated in the presence of RAPA suppressed colitis as naturally-occurring Treg, while CD4^+ T cells cultured in the absence of RAPA failed to ameliorate intestinal inflammation. Therefore, RAPA induced Treg, which are capable of suppressing colitis.These results indicate that Treg can be expanded or induced in vitro and transfer of induced Treg may be efficacious for the treatment of inflammatory bowel diseases.

CD4^+CD25^+调节性T细胞（Treg）具有广泛的免疫调节活性，可调节动物模型中的自身免疫性疾病，包括炎症性肠病。针对Treg治疗溃疡性结肠炎（UC）的临床应用，我们研究了Treg是否可以在体外扩增或诱导。我们用带有抗 CD3/抗 CD28 的珠子和 IL-2 刺激从 UC 患者的白细胞去除术产物中分离出的 Tregs。培养10天后，Treg可增加18倍。培养的 Treg 维持 Treg 特异性标记物 FOXP3 的表达，并在共培养时显着抑制 CD4^+ T 细胞增殖。因此，Treg可以在体外扩增。接下来，我们测试了Treg是否可以在体外被诱导。CD4^+CD255non-Treg在TGF-β1存在下刺激和培养。培养的细胞表达 FOXP3 并在共培养时抑制 CD4^+ T 细胞增殖。因此，甚至可以在体外从非Treg诱导Treg。然后，我们研究诱导的Treg是否能够抑制肠道炎症。我们在雷帕霉素 (RAPA) 存在的情况下，用抗 CD3/抗 CD28 和 IL-2 刺激 CD4^+ T 细胞诱导 Treg。当在 RAPA 存在下刺激 CD4^+ T 细胞时，在 21 天培养结束时，17% 为 CD25^+ FoxP3^+。相反，没有 RAPA 刺激的细胞很少表达 CD25 和 FoxP3。 CB-17 Scid 小鼠被转移 CD4^+CD62L^+CD25^-T 细胞，导致慢性持续性结肠炎。在 RAPA 存在的情况下刺激的 CD4^+ T 细胞作为天然 Treg 抑制结肠炎，而在不存在 RAPA 的情况下培养的 CD4^+ T 细胞未能改善肠道炎症。因此，RAPA诱导的Treg能够抑制结肠炎。这些结果表明Treg可以在体外扩增或诱导，并且诱导的Treg的转移可能对治疗炎症性肠病有效。

项目成果

The development of leukapheresis/regulatory T cell transfer therapy for the treatment of ulcerative colitis : the establishment of aseptic cell isolation protocol

白细胞分离术/调节性T细胞转移疗法治疗溃疡性结肠炎的进展：无菌细胞分离方案的建立

• 发表时间: 2007
• 作者: Sumida;Y.;Nakamura;K.;Kanayama;K.;Takahashi;M.;Mizutani;T.;Honda;K.;Higuchi;N.;Ogino;H.;Murao;H.;Taki;K.;Itaba;S.;Akiho;H.;Takayanagi;R
• 通讯作者: R

潰瘍性大腸炎に対する血球成分除去制御性T細胞分離・移入療法の開発:無菌的細胞分離法の確立

溃疡性结肠炎血细胞耗竭调节性T细胞分离和转移治疗的发展：无菌细胞分离方法的建立

• 发表时间: 2007
• 作者: 隅田 頼信;他
• 通讯作者: 他

An inverse correlation of human peripheral blood regulatory T cell frequency with the disease actlvity of ulcerative colitis

人外周血调节性T细胞频率与溃疡性结肠炎疾病活动性的负相关

• 发表时间: 2006
• 期刊: Digestive Diseases and Sciences 51
• 作者: Takahashi;et. al.
• 通讯作者: et. al.

An inverse correlation of human peripheral blood regulatory T cell frequency with the disease activity of ulcerative colitis

• DOI: 10.1007/s10620-006-3191-2
• 发表时间: 2006-04-01
• 期刊: DIGESTIVE DISEASES AND SCIENCES
• 影响因子: 3.1
• 作者: Takahashi, M;Nakamura, K;Nawata, H
• 通讯作者: Nawata, H

制御性T細胞をどう治療に生かすか?

我们如何利用调节性T细胞进行治疗？

• 发表时间: 2006
• 期刊: 炎症と免疫 14
• 作者: Takahashi;M.;Nakamura;K.;Honda;K.;Kitamura;Y.;Mizutani;T.;Araki;Y.;Kabemura;T.;Chijiiwa;Y.;Harada;N.;Nawata;H;Takahashi et al.;中村和彦
• 通讯作者: 中村和彦