Identification of the genes differentially expressed in human dendritic cell subsets by cDNA subtraction and microarray analysis
Identification of the genes differentially expressed in human dendritic cell subsets by cDNA subtraction and microarray analysis
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DOI：
10.1182/blood.v100.5.1742.h81702001742_1742_1754
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发表时间：
2002-09-01
期刊：
影响因子：
20.3
通讯作者：
Bae, YS
中科院分区：
文献类型：
--
作者：
Ahn, JH;Lee, Y;Bae, YS
最近的研究树突状细胞（DC）相关基因已进行使用单核细胞衍生的DC（MoDC）在不同的成熟阶段。为了揭示DC相关的新基因及其在不同DC亚群中的表达谱，我们构建了CD 1a（+）、CD 14（+）和CD 11 c（-）DC的消减DC cDNA文库，通过消减T细胞、B细胞和单核细胞共有的基因，然后借助微阵列技术筛选文库。选择在微阵列分析中对DC表现出显着特异性的基因，并通过半定量逆转录酶-聚合酶链反应进行确认。我们的研究发现：（1）在髓样DC中高表达的基因是那些参与抗原摄取/加工/呈递、细胞变态或趋化性的基因。(2)以前在MoDC中鉴定的大多数基因，如TARC、铁蛋白L链、溶酶体酸性脂肪酶、α-和β-微管蛋白、骨桥蛋白（Eta-1）等，在CD 11 c（-）DC中不显著表达，无论其成熟状态如何。另一方面，特异性转录因子和MHC II类分子，如干扰素调节因子-4（IRF 4）和HLA-DR，在两种DC亚群中表达相似。(3)CD 14（+）DCs保留了组织DCs的独特特征，如“无CCR 7但有更多CCR 1”和“无TARC但有丰富MCP 1和Eta-1”的基因表达谱所证明。（4）免疫球蛋白（IG）超家族Z391 g、CD 20样前体、糖蛋白NMB（GPNMB）、转化生长因子β（TGF-β）诱导蛋白（TGFBI）、髓样DAP 12相关凝集素（MDL-1）和6个新基因的基因被新鉴定为与DC亚群的表型表达相关。这些鉴定为进一步研究DC亚群的功能提供了重要的分子信息。
Recent studies on dendritic cell (DC)associated genes have been performed using monocyte-derived DCs (MoDCs) in different maturation stages. In our approach, to uncover the novel DC-associated genes and their expression profiles among the different DC subsets, we constructed a subtracted DC-cDNA library from CD1a(+), CD14(+), and CD11c(-) DCs by subtracting the genes shared with T cells, B cells, and monocytes, and we then screened the libraries with the aid of microarray technique. The genes showing remarkable specificity to DCs in the microarray analysis were selected and confirmed by semi-quantitative reverse transcriptase-polymerase chain reaction. Our investigations revealed the following: (1) Genes highly expressed in myeloid DCs are those involved in antigen uptake/processing/presentation, cell metamorphosis, or chemotaxis. (2) Most of the genes previously identified in MoDCs, such as TARC, ferritin L-chain, lysosomal acid lipase, alpha-and beta-tubulin, osteopontin (Eta-1), and others, are not markedly expressed in CD11c(-) DCs regardless of their maturation status. On the other hand, specific transcription factors and MHC class II molecules, such as interferon regulatory factor-4 (IRF4) and HLA-DR, are similarly expressed in both DC subsets. (3) CD14(+) DCs retain unique features of tissue DCs, as evidenced by the gene expression profile of "no CCR7 but more CCR1" and "no TARC but abundant MCP1 and Eta-1." (4) The genes for immunoglobulin (Ig) superfamily Z391g, CD20-like precursor, glycoprotein NMB (GPNMB), transforming growth factorbeta (TGF-beta)-induced protein (TGFBI), myeloid DAP12-associated lectin (MDL-1), and 6 novel genes are newly identified as being associated with the phenotypic expression of the DC subsets. These identifications provide important molecular information for further functional studies of the DC subsets.