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Scanning. 2016 Jul;38(4):365-75. doi: 10.1002/sca.21279. Epub 2015 Oct 28.

Detection of CD28/CD86 co-stimulatory molecules and surface properties of T and dendritic cells: An AFM study.

Huang X1,2, Guo H1,2, Wang C1,2, Mu J1,2, Zhang H1,2, Liang Z1,2, Cai J3, Zhou C1,2.

Author information

1
Department of Materials Science and Engineering, Jinan University, Guangzhou, China.
2
Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou, China.
3
Department of Chemistry, Jinan University, Guangzhou, China.

Abstract

Although the importance of B7/CD28 co-stimulation has been widely studied, little is known about their nano-spatial localization and their corresponding cells' biophysical and biomechanical properties. Here, we investigated the morphological, biophysical, and biomechanical properties of T cells and dendritic cells (DCs) by atomic force microscopy (AFM) and force curves. The nano-spatial distribution of CD28 and CD86 antigen on T cells and DCs was detected by CD86 or CD28 antibody-functionalized AFM tip. Single-molecule force spectroscopy (SMFS)-based force volumes and quantum dots (QDs)-based fluorescence imaging demonstrated that the co-stimulatory molecules were not randomly distributed over the cells' surface, but more than 80% of CD28 and CD86 molecules appeared to be expressed as 100-200 nm nanoclusters and polarize dominantly in the peak of the cell membrane fluctuations. AFM imaging and quantitative analysis showed that the roughness of mature DCs (mDCs) was higher than that of immature DCs (iDCs). The adhesion force distribution of iDCs and mDCs was heterogeneous while the elasticity distribution was homogeneous locally. In addition, mDCs had a fourfold increase of Young's modulus of iDCs, indicating the contribution of the actin cytoskeleton to the elastic properties of the cells. Taken together, the nano-cluster distribution of CD28 and CD86, the rough mDCs surface, the higher adhesion force and elasticity of mDCs may facilitate to the occurrence of B7/CD28 co-stimulation signals and the formation of immune synapse. These nanoscale findings provide new insights into the antigen-presenting function of DCs, the T cell activation and ultimate immune response. SCANNING 38:365-375, 2016.

KEYWORDS:

CD28/CD86; T cells; dendritic cells; single-molecule force spectroscopy (SMFS)

PMID:
26507362
DOI:
10.1002/sca.21279
[Indexed for MEDLINE]

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