Selective and efficient generation of functional Batf3-dependent CD103+ dendritic cells from mouse bone marrow
Selective and efficient generation of functional Batf3-dependent CD103+ dendritic cells from mouse bone marrow
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DOI：
10.1182/blood-2013-12-545772
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发表时间：
2014-11-13
期刊：
影响因子：
20.3
通讯作者：
Sparwasser, Tim
中科院分区：
文献类型：
--
作者：
Mayer, Christian Thomas;Ghorbani, Peyman;Sparwasser, Tim
FMS样酪氨酸激酶3配体（FLT 3L）依赖性树突状细胞（DC）的多个亚群控制T细胞耐受性和免疫力。在小鼠中，Batf 3依赖性CD 103（+）DC有效地进入淋巴结并交叉呈递抗原，使这种保守的DC亚群成为耐受诱导或疫苗接种的有希望的靶标。然而，用目前的方法只能分离有限数量的CD 103（+）DC。已建立的骨髓培养方案有效地产生单核细胞衍生的DC或产生FLT 3L依赖性DC亚群的混合物。我们发现，CD 103（+）DC的发展需要延长的培养时间和FLT 3L和粒细胞巨噬细胞集落刺激因子（GM-CSF）的持续作用，这可以通过GM-CSF对DC前体和分化的CD 103（+）DC的双重作用来解释。因此，我们建立了一种新的方法来产生大量的CD 103（+）DC（iCD 103-DC），而其他DC亚群的存在有限。iCD 103-DC以Batf 3和Irf 8依赖性方式发育，表达CD 8 α/CD 103 DC基因特征，交叉呈递细胞相关抗原，并对TLR 3刺激应答。因此，iCD 103-DC反映了组织CD 103（+）DC的关键特征。重要的是，iCD 103-DC在成熟后表达高水平的CCR 7，并且比经典的单核细胞来源的DC更有效地迁移到淋巴结。最后，iCD 103-DCs在体内诱导T细胞介导的保护性免疫。我们的研究提供了对CD 103（+）DC发育和功能的见解。
Multiple subsets of FMS-like tyrosine kinase 3 ligand (FLT3L)-dependent dendritic cells (DCs) control T-cell tolerance and immunity. In mice, Batf3-dependent CD103(+) DCs efficiently enter lymph nodes and cross-present antigens, rendering this conserved DC subset a promising target for tolerance induction or vaccination. However, only limited numbers of CD103(+) DCs can be isolated with current methods. Established bone marrow culture protocols efficiently generate monocyte-derived DCs or produce a mixture of FLT3L-dependent DC subsets. We show that CD103(+) DC development requires prolonged culture time and continuous action of both FLT3L and granulocyte macrophage colony-stimulating factor (GM-CSF), explained by a dual effect of GM-CSF on DC precursors and differentiating CD103(+) DCs. Accordingly, we established a novel method to generate large numbers of CD103(+) DCs (iCD103-DCs) with limited presence of other DC subsets. iCD103-DCs develop in a Batf3- and Irf8-dependent fashion, express a CD8 alpha/CD103 DC gene signature, cross-present cell-associated antigens, and respond to TLR3 stimulation. Thus, iCD103-DCs reflect key features of tissue CD103(+) DCs. Importantly, iCD103-DCs express high levels of CCR7 upon maturation and migrate to lymph nodes more efficiently than classical monocyte-derived DCs. Finally, iCD103-DCs induce T cell-mediated protective immunity in vivo. Our study provides insights into CD103(+) DC development and function.