In Vitro Generation of Human XCR1+ Dendritic Cells from CD34+ Hematopoietic Progenitors
In Vitro Generation of Human XCR1+ Dendritic Cells from CD34+ Hematopoietic Progenitors
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DOI：
10.1007/978-1-4939-3606-9_2
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发表时间：
2016-01-01
期刊：
影响因子：
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通讯作者：
Dalod, Marc
中科院分区：
文献类型：
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作者：
Balan, Sreekumar;Dalod, Marc
树突状细胞（Dendritic cells，DCs）是一种异质性的专职抗原呈递细胞，在免疫防御中起着关键作用。获得的关于人DC生物学的大多数信息来自于对体外从外周血CD14（+）单核细胞（MoDC）或从CD34（+）造血祖细胞产生的DC进行的研究。该领域的最新进展表明，这些类型的体外衍生的DC与天然存在于人类淋巴器官中的DC亚群在全局基因表达、感测不同类型的危险信号的专门化以及向不同功能调节T淋巴细胞的能力方面显著不同。目前正在努力更好地表征人类淋巴器官驻留DC亚群的生物学和功能，这是设计针对感染或癌症的基于DC的创新疫苗的重要步骤。然而，这条研究路线受到大多数组织中某些DC亚群的低频率，它们的脆弱性以及纯化所需程序的复杂性的阻碍。因此，需要稳健的程序，其允许在允许其遗传或药理学操作的条件下大规模体外产生人DC亚群，以破译其功能和其分子调控。人CD141（+）CLEC9A（+）XCR1（+）DC构成了一个非常有趣的DC亚群，用于设计针对细胞内病原体感染或癌症的免疫治疗，因为这些细胞类似于小鼠专业交叉呈递CD8 α（+）Clec9a（+）Xcr1（+）DC。人XCR1（+）DC确实已经被几个团队报道为比其他人DC亚群更有效地交叉呈递，特别是细胞相关抗原的交叉呈递，但也是可溶性抗原的交叉呈递，特别是当递送到晚期内体或溶酶体中时。然而，人XCR1（+）DC是最稀有的，可能是最脆弱的人DC亚群，因此最难以离体研究。在这里，我们描述了一种允许同时体外产生人MoDC和XCR 1（+）DC的方案，这无疑将非常有用，以更好地表征人XCR 1（+）DC的功能特化并鉴定其分子基础。
Dendritic cells (DCs) are a heterogeneous population of professional antigen-presenting cells which play a key role in orchestrating immune defenses. Most of the information gained on human DC biology was derived from studies conducted with DCs generated in vitro from peripheral blood CD14(+) monocytes (MoDCs) or from CD34(+) hematopoietic progenitors. Recent advances in the field revealed that these types of in vitro-derived DCs strikingly differ from the DC subsets that are naturally present in human lymphoid organs, in terms of global gene expression, of specialization in the sensing of different types of danger signals, and of the ability to polarize T lymphocytes toward different functions. Major efforts are being made to better characterize the biology and the functions of lymphoid organ-resident DC subsets in humans, as an essential step for designing innovative DC-based vaccines against infections or cancers. However, this line of research is hampered by the low frequency of certain DC subsets in most tissues, their fragility, and the complexity of the procedures necessary for their purification. Hence, there is a need for robust procedures allowing large-scale in vitro generation of human DC subsets, under conditions allowing their genetic or pharmacological manipulation, to decipher their functions and their molecular regulation. Human CD141(+)CLEC9A(+)XCR1(+) DCs constitute a very interesting DC subset for the design of immunotherapeutic treatments against infections by intracellular pathogens or against cancer, because these cells resemble mouse professional cross-presenting CD8 alpha(+)Clec9a(+)Xcr1(+) DCs. Human XCR1(+) DCs have indeed been reported by several teams to be more efficient than other human DC subsets for cross-presentation, in particular of cell-associated antigens but also of soluble antigens especially when delivered into late endosomes or lysosomes. However, human XCR1(+) DCs are the rarest and perhaps the most fragile of the human DC subsets and hence the most difficult to study ex vivo. Here, we describe a protocol allowing simultaneous in vitro generation of human MoDCs and XCR1(+) DCs, which will undoubtedly be extremely useful to better characterize the functional specialization of human XCR1(+) DCs and to identify its molecular bases.