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）潜在的广泛的免疫调节活性范围，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在治疗溃疡性结肠炎（UC）的临床应用中，我们研究了Treg是否可以在体外扩张或诱导。我们刺激了从抗CD3/抗CD28的抗CD28珠和IL-2的UC患者中分离出的Treg。 Treg可以在10天的培养物中增加18次。培养的Treg保持了Treg特异性标记物FOXP3的表达，并在共培养后显着抑制了CD4^+ T细胞增殖。这是可以在体外扩展的Treg。我们测试了在TGF-β1存在下是否可以在体外CD4^+CD255non-Treg中诱导Treg。培养的细胞表达Foxp3并在共培养后抑制了CD4^+ T细胞增殖。因此，甚至可以在体外非特雷格诱导treg。然后，我们研究诱导的Treg是否能够抑制肠道注射。我们在存在雷帕霉素（RAPA）的情况下用抗CD3/抗CD28和IL-2刺激的CD4^+ T细胞诱导Treg。当CD4^+ T细胞在RAPA存在下刺激时，在21天培养结束时，17％为CD25^+ Foxp3^+。相比之下，很少有没有RAPA表达CD25和FOXP3的细胞。用CD4^+CD62L^+CD25^-T细胞转移CB-17 SCID小鼠，该细胞发展出慢性持续性结肠炎。在Rapa存在下刺激的CD4^+ T细胞的共转移是自然呈现的Treg，而在没有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可以在体外扩展或诱导，并且诱导的Treg转移可能有效地治疗炎症性肠病。

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 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

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.

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