Dynamic Regulation of Notch 1 and Notch 2 Surface Expression during T Cell Development and Activation Revealed by Novel Monoclonal Antibodies
Dynamic Regulation of Notch 1 and Notch 2 Surface Expression during T Cell Development and Activation Revealed by Novel Monoclonal Antibodies
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DOI：
10.4049/jimmunol.0902432
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发表时间：
2009-12-01
影响因子：
4.4
通讯作者：
MacDonald, H. Robson
中科院分区：
文献类型：
--
作者：
Fiorini, Emma;Merck, Estelle;MacDonald, H. Robson
众所周知，Notch信号传导在T细胞发育和活化的多个阶段起关键作用。然而，与T细胞中Notch信号传导相关的细胞和分子事件的详细分析受到缺乏可以明确测量细胞表面Notch受体表达的试剂的阻碍。使用针对Notch 1和Notch 2细胞外结构域的新型大鼠单克隆抗体，我们发现Notch 1已经在骨髓中常见的淋巴前体细胞上高度表达，并且在CD 4（-）CD 8（-）胸腺细胞的胸腺内成熟过程中保持高水平。Notch 1在CD 4（+）CD 8（+）和成熟CD 4（+）或CD 8（+）胸腺阶段逐渐下调，并在外周T细胞上以低水平表达。胸腺冷冻切片的免疫荧光染色进一步揭示了Notch 1（+）CD 25（-）细胞邻近胸腺囊的定位。Notch 1在体外用抗CD 3单克隆抗体激活或体内用淋巴细胞性脉络膜脑膜炎病毒或利什曼原虫感染后在外周T细胞上上调。与Notch 1相反，Notch 2在常见的淋巴前体和CD 117（+）早期胸腺内亚群上以中等水平表达，但在T细胞发育的后续阶段完全消失。然而，在抗CD 3刺激后，在外周T细胞上也观察到Notch 2的瞬时上调。总的来说，我们的新型mAb揭示了在T细胞发育和活化期间Notch 1和Notch 2表面表达的动态调节。此外，它们为将来分析包括造血系统在内的各种组织中的Notch受体提供了重要资源。免疫学杂志，2009，183：7212-7222.
It is well established that Notch signaling plays a critical role at multiple stages of T cell development and activation. However, detailed analysis of the cellular and molecular events associated with Notch signaling in T cells is hampered by the lack of reagents that can unambiguously measure cell surface Notch receptor expression. Using novel rat mAbs directed against the extracellular domains of Notch1 and Notch2, we find that Notch1 is already highly expressed on common lymphoid precursors in the bone marrow and remains at high levels during intrathymic maturation of CD4(-)CD8(-) thymocytes. Notch1 is progressively down-regulated at the CD4(+)CD8(+) and mature CD4(+) or CD8(+) thymic stages and is expressed at low levels on peripheral T cells. Immunofluorescence staining of thymus cryosections further revealed a localization of Notch1(+)CD25(-) cells adjacent to the thymus capsule. Notch1 was up-regulated on peripheral T cells following activation in vitro with anti-CD3 mAbs or infection in vivo with lymphocytic chorio-meningitis virus or Leishmania major. In contrast to Notch1, Notch2 was expressed at intermediate levels on common lymphoid precursors and CD117(+) early intrathymic subsets, but disappeared completely at subsequent stages of T cell development. However, transient up-regulation of Notch2 was also observed on peripheral T cells following anti-CD3 stimulation. Collectively our novel mAbs reveal a dynamic regulation of Notch1 and Notch2 surface expression during T cell development and activation. Furthermore they provide an important resource for future ana lysis of Notch receptors in various tissues including the hematopoietic system. The Journal of Immunology, 2009, 183: 7212-7222.