Introduction of Human Flt3-L and GM-CSF into Humanized Mice Enhances the Reconstitution and Maturation of Myeloid Dendritic Cells and the Development of Foxp3(+)CD4(+) T Cells.
Introduction of Human Flt3-L and GM-CSF into Humanized Mice Enhances the Reconstitution and Maturation of Myeloid Dendritic Cells and the Development of Foxp3(+)CD4(+) T Cells.
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DOI：
10.3389/fimmu.2018.01042
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发表时间：
2018
影响因子：
7.3
通讯作者：
Terahara K
中科院分区：
文献类型：
--
作者：
Iwabuchi R;Ikeno S;Kobayashi-Ishihara M;Takeyama H;Ato M;Tsunetsugu-Yokota Y;Terahara K
fms相关酪氨酸激酶3配体（Flt 3-L）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）被认为是体内树突状细胞（DC）发育的重要调节因子。然而，Flt 3-L和GM-CSF对人DC的联合作用尚未在体内评估。因此，在这项研究中，我们的目的是使用人源化小鼠模型进行评估。通过将来自人脐带血的造血干细胞移植到新生NOJ小鼠中构建人源化非肥胖糖尿病/SCID/Jak 3 null（hNOJ）小鼠，并使用编码人Flt 3-L和GM-CSF的质粒通过流体动力学注射介导的基因递送进行体内转染（IVT）。IVT后，在hNOJ小鼠中成功诱导Flt 3-L和GM-CSF。在IVT后10天，我们发现，在脾脏中，Flt 3-L和GM-CSF治疗增强了两种髓样DC亚群的重建，除了表达CD 1c和/或CD 141的CD 14+单核细胞样细胞外，CD 14 − CD 1c+常规DC（cDC）和CD 14 − CD 141 + cDC。单独的GM-CSF对这些骨髓细胞群的重建作用较小。相比之下，没有细胞因子处理增强CD 123+浆细胞样DC（pDC）重建。无论重建水平如何，三种细胞群（CD 1c+髓样细胞、CD 141+髓样细胞和pDC）可以通过用细胞因子处理而成熟，就上调CD 40、CD 80、CD 86和CD 184/CXCR 4以及下调CD 195/CCR 5而言。特别是，GM-CSF在所有这些细胞群中促进了CD 80的上调。有趣的是，我们进一步观察到，在GM-CSF存在下，脾CD 4 + T细胞内的Foxp 3+细胞显著增加。Foxp 3 + T细胞可以分为两个亚群，CD 45 RA − Foxp 3 hi和CD 45 RA − Foxp 3lo T细胞。而CD 45 RA − Foxp 3 hi T细胞仅在单独用GM-CSF处理后增加，CD 45 RA − Foxp 3lo T细胞仅在用Flt 3-L和GM-CSF处理后增加。单独用Flt 3-L处理对Foxp 3 + T细胞的数量没有影响。相关性分析表明，这些Foxp 3+亚群的发育与DC（-样）细胞的成熟状态有关。综上所述，本研究为研究Flt 3-L和GM-CSF对人DC和调节性T细胞的体内作用提供了平台。
Two cytokines, fms-related tyrosine kinase 3 ligand (Flt3-L) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are considered to be the essential regulators of dendritic cell (DC) development in vivo. However, the combined effect of Flt3-L and GM-CSF on human DCs has not been evaluated in vivo. In this study, we, therefore, aimed at evaluating this using a humanized mouse model. Humanized non-obese diabetic/SCID/Jak3null (hNOJ) mice were constructed by transplanting hematopoietic stem cells from human umbilical cord blood into newborn NOJ mice, and in vivo transfection (IVT) was performed by hydrodynamic injection-mediated gene delivery using plasmids encoding human Flt3-L and GM-CSF. Following IVT, Flt3-L and GM-CSF were successfully induced in hNOJ mice. At 10 days post-IVT, we found, in the spleen, that treatment with both Flt3-L and GM-CSF enhanced the reconstitution of two myeloid DC subsets, CD14−CD1c+ conventional DCs (cDCs) and CD14−CD141+ cDCs, in addition to CD14+ monocyte-like cells expressing CD1c and/or CD141. GM-CSF alone had less effect on the reconstitution of these myeloid cell populations. By contrast, none of the cytokine treatments enhanced CD123+ plasmacytoid DC (pDC) reconstitution. Regardless of the reconstitution levels, three cell populations (CD1c+ myeloid cells, CD141+ myeloid cells, and pDCs) could be matured by treatment with cytokines, in terms of upregulation of CD40, CD80, CD86, and CD184/CXCR4 and downregulation of CD195/CCR5. In particular, GM-CSF contributed to upregulation of CD80 in all these cell populations. Interestingly, we further observed that Foxp3+ cells within splenic CD4+ T cells were significantly increased in the presence of GM-CSF. Foxp3+ T cells could be subdivided into two subpopulations, CD45RA−Foxp3hi and CD45RA−Foxp3lo T cells. Whereas CD45RA−Foxp3hi T cells were increased only after treatment with GM-CSF alone, CD45RA−Foxp3lo T cells were increased only after treatment with both Flt3-L and GM-CSF. Treatment with Flt3-L alone had no effect on the number of Foxp3+ T cells. The correlation analysis demonstrated that the development of these Foxp3+ subpopulations was associated with the maturation status of DC(-like) cells. Taken together, this study provides a platform for studying the in vivo effect of Flt3-L and GM-CSF on human DCs and regulatory T cells.