IL-3-dependent murine mast cells undergo apoptosis on removal of IL-3. Prevention of apoptosis by c-kit ligand.
IL-3-dependent murine mast cells undergo apoptosis on removal of IL-3. Prevention of apoptosis by c-kit ligand.
复制标题
IL-3依赖性小鼠肥大细胞在去除IL-3后发生凋亡。
DOI：
10.4049/jimmunol.151.7.3775
复制
发表时间：
1993
影响因子：
4.4
通讯作者：
D. Metcalfe
中科院分区：
文献类型：
--
作者：
Y. Mekori;C. Oh;D. Metcalfe
肥大细胞的增殖和成熟受IL-3和c-kit配体干细胞因子(SCF)两种主要细胞因子的调节。然而，很少有人知道这两个过程是如何负调控的，从而知道肥大细胞数量在正常或病理过程中是如何控制的。在这项研究中，我们假设依赖于IL-3的肥大细胞在去除IL-3后会经历程序性细胞死亡(凋亡)，正如其他依赖生长因子的造血细胞所显示的那样。用光镜、吖啶橙荧光染色、流式细胞仪和DNA电泳法分析细胞凋亡的变化。我们可以证明，无论是从原代培养的骨髓来源的肥大细胞培养物中去除IL-3，还是从生长因子依赖的肥大细胞系MCP5中去除IL-3，都会导致细胞发生典型的凋亡变化，包括染色质浓缩、核碎裂、细胞空泡化，流式细胞仪检测显示典型的碘化丙啶或Hoechst 33342摄取模式，以及典型的200bp的DNA切割梯形模式。SCF可以阻止这些事件的发生，这一作用部分是由酪氨酸激酶介导的，因为酪氨酸激酶抑制剂赫比霉素抑制了SCF防止IL-3剥夺的细胞凋亡的作用。用从纯合子的W/WV小鼠获得的抗c-kit单抗和IL-3依赖的BMCMC对c-kit受体编码的w基因进行突变，也可以证明SCF通过c-kit发挥作用。地塞米松和环孢菌素A均不能抑制SCF的“救援”作用，提示“救援”是由SCF介导的，而不是通过其他细胞因子的诱导。因此，依赖于IL-3的肥大细胞在去除IL-3后发生凋亡，这一事件被SCF通过其配体c-kit添加而被阻止，从而表明在生理条件下这些主要的肥大细胞生长因子如何协同作用来调节肥大细胞的数量。
It is well established that mast cell proliferation and maturation are regulated by two principle cytokines, IL-3 and the c-kit ligand stem cell factor (SCF). Little is known, however, how these two processes are negatively regulated and thus, how mast cell number is controlled in normal or pathologic processes. In this study we hypothesized that IL-3-dependent mast cells would undergo programmed cell death (apoptosis) on removal of IL-3 as was shown with other growth factor-dependent hemopoietic cells. Apoptotic changes were analyzed using light microscopy, fluorescent staining with acridine orange, flow cytometric analysis, and DNA electrophoresis. We could demonstrate that elimination of IL-3 from either primary bone marrow-derived cultured mast cell cultures (BMCMC) or from the growth factor-dependent mast cell line MCP5 resulted in the characteristic changes of apoptosis including condensed chromatin, fragmented nuclei, cellular vacuolization, typical pattern of propidium iodide or Hoechst 33342 uptake by flow cytometry, and the characteristic 200 bp "ladder" pattern of DNA cleavage. These events were prevented by SCF, an action that was in part mediated by tyrosine kinases, in that the tyrosine kinase inhibitor herbimycin inhibited the action of SCF in preventing apoptosis in IL-3-deprived cells. By using anti c-kit mAb and IL-3-dependent BMCMC obtained from W/Wv mice homozygous for mutation at the w locus that encodes the c-kit receptor, we could also show that SCF exerted its effect via c-kit. Neither dexamethasone nor cyclosporin A inhibited the "rescue" effect of SCF, suggesting that "rescue" was mediated by SCF and not through the induction of other cytokines. Thus, IL-3-dependent mast cells undergo apoptosis on removal of IL-3, an event that is prevented by the addition of SCF through its ligand c-kit, thus demonstrating how these principle mast cell growth factors may act in concert to regulate mast cell number under physiologic conditions.