Isolation and characterization of human peripheral blood dendritic cells.
Isolation and characterization of human peripheral blood dendritic cells.
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DOI：
10.4049/jimmunol.150.3.821
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发表时间：
1993-02
影响因子：
4.4
通讯作者：
Ranjeny Thomas;Laurie S. Davis;P. E. Lipsky
中科院分区：
文献类型：
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作者：
Ranjeny Thomas;Laurie S. Davis;P. E. Lipsky
树突状细胞（DC）是有效的抗原提呈细胞，可从培养的外周血非T细胞中纯化出来。由于尚未发现特异性的细胞表面标志物，DC的谱系仍不明确。这些研究的目的是确定能够产生功能性人DC的循环血细胞。当用标准技术从经L - 亮氨酰 - L - 亮氨酸甲酯处理的非T细胞中纯化时，DC得到富集，已知该物质对单核细胞和细胞毒性细胞有毒性。为了确定单核细胞或B细胞是否能产生DC，将新鲜的非T细胞分选成CD14⁺单核细胞、CD19⁺ B细胞以及CD14⁻和CD19⁻细胞。尽管培养的非贴壁CD14⁻或CD19⁻细胞的抗原提呈细胞功能有一定程度的富集，但在通过不表达CD14、CD19、CD3和CD16进行分选的细胞群中，发现具有树突状形态和强大抗原提呈细胞功能的细胞有显著富集。超过90%的CD14⁻CD19⁻CD16⁻CD3⁻分选细胞以及对照DC表达髓系标志物CD13和CD33。因此，根据这些髓系标志物的表达对新鲜的非T细胞进行分选。与CD33⁻CD14⁻ B细胞相比，一些CD33⁺细胞微弱表达CD14。然而，它们很容易与强烈表达CD14的单核细胞区分开来。CD33⁺CD14dim细胞形成树突状突起，并且比对照DC、CD33⁺CD14⁺或CD33⁻CD14⁻细胞是更有效的抗原提呈细胞。尽管新鲜分离的CD33⁺CD14dim DC表达一些也由CD14⁺单核细胞表达的细胞表面分子，但与单核细胞相比，它们的溶酶体酶水平较低，且缺乏FcR介导的吞噬作用。CD33⁺CD14dim细胞的形态和表型分化在培养6到36小时内发生。然而，CD33⁺CD14dim细胞无需长时间预培养以形成树突状形态就能有效地作为抗原提呈细胞发挥作用。这些数据表明，人血液中的DC源自表达髓系谱系标志物CD13和CD33的前体细胞，但在功能上与经典的CD14⁺单核细胞不同。
Dendritic cells (DC) are potent APC that can be purified from cultured peripheral blood non-T cells. Because no specific cell surface marker has been found, the lineage of DC remains obscure. The purpose of these studies was to determine the circulating blood cells that could give rise to functional human DC. DC were enriched when purified by standard techniques from non-T cells that were treated with L-leucyl-L-leucine methyl ester, known to be toxic to monocytes and cytolytic cells. To determine whether monocytes or B cells could give rise to DC, fresh non-T cells were sorted into CD14+ monocytes, CD19+ B cells, and CD14- and CD19- cells. Although there was some enrichment for APC function by cultured nonadherent CD14- or CD19- cells, a marked enrichment for cells with dendritic morphology and potent APC function was found in the population that was sorted by the absence of expression of CD14, CD19, CD3, and CD16. More than 90% of the CD14-CD19-CD16-CD3- sorted cells, and of control DC, expressed the myeloid markers CD13 and CD33. Therefore, fresh non-T cells were sorted based on the expression of these myeloid markers. In comparison with CD33-CD14- B cells, some of the CD33+ cells expressed CD14 dimly. However, they were easily distinguished from monocytes, which intensely expressed CD14. CD33+CD14dim cells developed dendritic processes and were more potent APC than control DC, CD33+CD14+, or CD33-CD14- cells. Although freshly isolated CD33+CD14dim DC expressed a number of cell surface molecules also expressed by CD14+ monocytes, they demonstrated lower levels of lysosomal enzymes and a lack of FcR-mediated phagocytosis in comparison with monocytes. Differentiation of morphology and phenotype of CD33+CD14dim cells occurred within 6 to 36 h in culture. However, the CD33+CD14dim cells could effectively function as APC without prolonged preincubation to develop dendritic morphology. These data indicate that human blood DC arise from precursors that express the myeloid lineage markers CD13 and CD33, but are functionally distinct from classic CD14+ monocytes.