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- Title
AFM- and NSOM-based force spectroscopy and distribution analysis of CD69 molecules on human CD4<sup>+</sup> T cell membrane.
- Authors
Hu, Mingqian; Chen, Jianan; Wang, Jiongkun; Wang, Xiaoping; Ma, Shuyuan; Cai, Jiye; Chen, Crystal Y.; Chen, Zheng W.
- Abstract
Although CD69 is well known as an early T cell-activation marker, the possibility that CD69 are distributed as nano-structures on membrane for immune regulation during T cell activation has not been tested. In this study, nanoscale features of CD69 expression on activated T cells were determined using the atomic force microscopy (AFM) topographic and force-binding nanotechnology as well as near-field scanning optical microscopy (NSOM)-/fluorescence quantum dot (QD)-based nanosacle imaging. Unstimulated CD4+ T cells showed neglectable numbers of membrane CD69 spots binding to the CD69 Ab-functinalized AFM tip, and no detectable QD-bound CD69 as examined by NSOM/QD-based imaging. In contrast, Phytohemagglutinin (PHA)-activated CD4+ T cells expressed CD69, and displayed many force-binding spots binding to the CD69 Ab-functionalized AFM tip on about 45% of cell membrane, with mean binding-rupture forces 276 ± 71 pN. Most CD69 molecules appeared to be expressed as 100-200 nm nanoclusters on the membrane of PHA-activated CD4+ T cells. Meanwhile, NSOM/QD-based nanoscale imaging showed that CD69 were non-uniformly distributed as 80-200 nm nanoclusters on cell-membrane of PHA-activated CD4+ T cells. This study represents the first demonstration of the nano-biology of CD69 expression during T cell activation. Copyright © 2009 John Wiley & Sons, Ltd.
- Publication
Journal of Molecular Recognition, 2009, Vol 22, Issue 6, p516
- ISSN
0952-3499
- Publication type
Article
- DOI
10.1002/jmr.976