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由CD8α(-)脾树突状细胞差异表达的C型凝集素超家族蛋白的分子克隆

Molecular cloning of a C-type lectin superfamily protein differentially expressed by CD8alpha(-) splenic dendritic cells.

作者信息

Caminschi I, Lucas K M, O'Keeffe M A, Hochrein H, Laâbi Y, Brodnicki T C, Lew A M, Shortman K, Wright M D

机构信息

The Walter and Eliza Hall Institute of Medical Research, PO Royal Melbourne Hospital, Victoria 3050, Melbourne, Australia.

出版信息

Mol Immunol. 2001 Sep;38(5):365-73. doi: 10.1016/s0161-5890(01)00067-0.

Abstract

Dendritic cells (DC) are potent antigen presenting cells that activate naive T cells. It is becoming increasingly clear that DC are not a homogeneous cell population, but comprise different subpopulations that differ in ontogeny and function. To further the molecular characterisation of DC, we screened for genes that were differentially expressed amongst DC subsets and could therefore give insight into their varying biological functions. Using Representational Difference Analysis (RDA) we identified a gene (CIRE) that is expressed at higher levels in the myeloid-related CD8alpha(-) DC than in the lymphoid-related CD8alpha(+) DC. CIRE is a 238 amino acid type II membrane protein, of approximately 33 kDa in size, whose extracellular region contains a C-type lectin domain. Northern blot analysis revealed that CIRE is almost exclusively expressed in DC and was not detected in organs such as heart, brain, kidney, liver, and thymus. T cells failed to express message for CIRE, whilst B cells expressed very low levels. These data here further substantiated by Northern blot analysis of 18 cell lines of various origins (myeloid, macrophage, B and T cell) where only one cell line, which was of myeloid origin and could give rise to DC, expressed mRNA for CIRE. Semi-quantitative RT-PCR suggested that CIRE is down-regulated upon activation. CIRE shares 57% identity with human DC-SIGN, a molecule that has been shown to be the ligand of ICAM-3 and that is also a receptor that binds HIV and facilitates trans-infection of T cells.

摘要

树突状细胞(DC)是激活初始T细胞的高效抗原呈递细胞。越来越清楚的是,DC不是一个同质的细胞群体,而是由在个体发生和功能上不同的亚群组成。为了进一步对DC进行分子特征分析,我们筛选了在DC亚群中差异表达的基因,因此可以深入了解它们不同的生物学功能。使用代表性差异分析(RDA),我们鉴定了一个基因(CIRE),它在髓样相关的CD8α(-)DC中比在淋巴样相关的CD8α(+)DC中表达水平更高。CIRE是一种238个氨基酸的II型膜蛋白,大小约为33 kDa,其细胞外区域包含一个C型凝集素结构域。Northern印迹分析显示,CIRE几乎只在DC中表达,在心脏、大脑、肾脏、肝脏和胸腺等器官中未检测到。T细胞未能表达CIRE的信息,而B细胞表达水平非常低。对18种不同来源(髓样、巨噬细胞、B和T细胞)的细胞系进行Northern印迹分析进一步证实了这些数据,其中只有一个髓样来源且可分化为DC的细胞系表达CIRE的mRNA。半定量RT-PCR表明,CIRE在激活后下调。CIRE与人类DC-SIGN有57%的同源性,DC-SIGN已被证明是ICAM-3的配体,也是一种结合HIV并促进T细胞转染的受体。

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