Nanomapping of CD1d–glycolipid complexes on THP1 cells by using simultaneous topography and recognition imaging
Nanomapping of CD1d–glycolipid complexes on THP1 cells by using simultaneous topography and recognition imaging
复制标题
使用同步形貌和识别成像对 THP1 细胞上的 CD1d-糖脂复合物进行纳米绘图
DOI：
10.1002/jmr.2282
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发表时间：
2013
影响因子：
2.7
通讯作者：
P. Hinterdorfer
中科院分区：
文献类型：
--
作者：
M. Duman;L. Chtcheglova;R. Zhu;B. Bozna;P. Polzella;V. Cerundolo;P. Hinterdorfer
CD1d 分子是一种单态主要组织相容性复合物 I 类样分子，向不变自然杀伤 T (iNKT) 细胞呈递不同类型的糖脂，在感染和肿瘤免疫以及调节自身免疫方面发挥重要作用。在这里，我们提出了同步形貌和识别成像（TREC）分析，以检测加载不同糖脂的 THP1 细胞上单个 CD1d 分子的密度、分布和定位。 TREC 使用带有生物素化 iNKT 细胞受体 (TCR) 功能的磁性涂层原子力显微镜尖端进行。识别图显示结合位点可见为黑点，这是由于 iNKT TCR 与 CD1d-糖脂复合物之间特异性结合期间振荡幅度减小所致。 THP1 细胞用三种不同的糖脂（α-GalCer、C20 和 OCH12）脉冲 4 小时和 16 小时。细胞膜上的 CD1d-α-GalCer 和 CD1d-C20:2 复合物形成了较小的微域，可达约 10 000nm2（尺寸面积），而 OCH12 负载的 CD1d 复合物则呈现出更大的簇，尺寸可达约 30 000nm2。此外，识别点的最小尺寸约为25nm，对应于单个CD1d结合位点。 TREC 在生理条件下以单分子分辨率成功揭示了 THP1 细胞上 CD1d-糖脂复合物的分布和定位。版权所有 © 2013 约翰·威利父子有限公司
CD1d molecule, a monomorphic major histocompatibility complex class I‐like molecule, presents different types of glycolipids to invariant natural killer T (iNKT) cells that play an important role in immunity to infection and tumors, as well as in regulating autoimmunity. Here, we present simultaneous topography and recognition imaging (TREC) analysis to detect density, distribution and localization of single CD1d molecules on THP1 cells that were loaded with different glycolipids. TREC was conducted using magnetically coated atomic force microscopy tips functionalized with a biotinylated iNKT cell receptor (TCR). The recognition map revealed binding sites visible as dark spots, resulting from oscillation amplitude reduction during specific binding between iNKT TCR and the CD1d–glycolipid complex. THP1 cells were pulsed with three different glycolipids (α‐GalCer, C20 and OCH12) for 4 and 16 hr. Whereas CD1d–α‐GalCer and CD1d–C20:2 complexes on cellular membrane formed smaller microdomains up to ~10 000 nm2 (dimension area), OCH12 loaded CD1d complexes presented larger clusters with a dimension up to ~30 000 nm2. Moreover, the smallest size of recognition spots was about 25 nm, corresponding to a single CD1d binding site. TREC successfully revealed the distribution and localization of CD1d–glycolipid complexes on THP1 cell with single molecule resolution under physiological conditions. Copyright © 2013 John Wiley & Sons, Ltd.
影响因子：
7.3
作者：
Keller CW;Freigang S;Lünemann JD
通讯作者：
Lünemann JD
DOI：
10.1073/pnas.0407488102
发表时间：
2005-03-01
影响因子：
11.1
作者：
Yu, KOA;Im, JS;Porcelli, SA
通讯作者：
Porcelli, SA