In-vitro generation and characterisation of murine CD4+CD25+ regulatory T cells with indirect allospecificity
In-vitro generation and characterisation of murine CD4+CD25+ regulatory T cells with indirect allospecificity
复制标题
DOI：
10.1016/j.intimp.2006.07.032
复制
发表时间：
2006-12-20
影响因子：
5.6
通讯作者：
Lechler, Robert I.
中科院分区：
文献类型：
--
作者：
Tsang, Julia;Jiang, Shuiping;Lechler, Robert I.
自然产生的CD4(+)CD25(+)调节性T细胞在预防自身免疫和诱导供体特异性移植耐受中发挥着关键作用。利用调节细胞进行潜在的过继细胞治疗由于缺乏抗原特异性且数量有限而受到阻碍。在这里，我们描述了对所有 K-d 肽具有抗原特异性的鼠 CD4(+)CD25(+) T 细胞的生成和扩增，作为过继细胞疗法的潜在试剂，以促进供体特异性移植耐受。使用用 K-d 肽脉冲的骨髓来源的自体树突状细胞，我们从 C56BL/6 小鼠中产生了纯化的 CD4(+)CD25(+) T 细胞的 T 细胞系。 T细胞系表达高水平的CD25和低水平的CD45RB和CD69。他们维持了 CD62L、GITR、CTLA-4 以及更重要的是 FoxP3 的表达。 CD4(+)CD25(+)T细胞系经TCR刺激后无反应性，几乎不产生IL-2、IFN-γ等细胞因子。重要的是，它们比新鲜分离的CD4(+)CD25(+) T细胞在抑制效应CD4(+) T细胞的增殖和细胞因子分泌方面更有效。此外，CD4(+)CD25(+) T细胞系可以扩增至大量细胞并在培养物中维持长达1年。 K-d特异性CD4(+)CD25(+) T细胞系对于设计在携带I完全错配BALB/c心脏的野生型B6小鼠中诱导心脏移植耐受的策略将具有无价的价值。 (c) 2006 Elsevier B.V. All Lights 保留。
Naturally arising CD4(+)CD25(+) regulatory T cells play a pivotal role in the prevention of autoimmunity and in the induction of donor-specific transplantation tolerance. Harnessing regulatory cells for potential adoptive cell therapy is hampered by their lack of antigen-specificity and their limited numbers. Here we describe the generation and expansion of murine CD4(+)CD25(+) T cells with antigen-specificity for all K-d peptide as potential reagents for adoptive cell therapy in promoting donor-specific transplantation tolerance. Using bone marrow-derived autologous dendritic cells Pulsed with the K-d peptide, we generated T cell lines front purified CD4(+)CD25(+) T cells from C56BL/6 mice. The T cell lines expressed high level of CD25 and low level of CD45RB and CD69. They maintained the expression of CD62L, GITR, CTLA-4 and more importantly FoxP3. The CD4(+)CD25(+) T cell lines were anergic after TCR stimulation and produced little cytokine such as IL-2 and IFN-gamma. Importantly, they were more potent than freshly isolated CD4(+)CD25(+) T cells in suppressing proliferation and cytokine secretion by effector CD4(+) T cells. Furthermore, the CD4(+)CD25(+) T cell lines Could be expanded to large cell numbers and maintained in culture up to I year. The K-d-specific CD4(+)CD25(+) T cell lines will be invaluable in devising a strategy for the induction of cardiac transplantation tolerance in wild-type B6 mice carrying I full mismatch BALB/c heart. (c) 2006 Elsevier B.V. All Lights reserved.