Single cell force spectroscopy of T cells recognizing a myelin-derived peptide on antigen presenting cells.
T-cell recognition of peptide-MHC complexes on APCs requires cell-cell interactions. The molecular events leading to T-cell activation have been extensively investigated, but the underlying physical binding forces between T-cells and APCs are largely unknown. We used single cell force spectroscopy for quantitation of interaction forces between T-cells and APCs presenting a tolerogenic peptide derived from myelin basic protein. When T-cells were brought into contact with peptide-loaded APCs, interaction forces increased with time from about 0.5nN after 10s interaction to about 15nN after 30min. In the absence of antigen, or when ICAM-1-negative APC was used, no increase in binding forces was observed. The temporal development of interaction forces correlated with the kinetics of immune synapse formation, as determined by LFA-1 and TCR enrichment at the interface of T-cell/APC conjugates using high throughput multispectral imaging flow cytometry. Together, these results suggest that ICAM-1/LFA-1 redistribution to the contact area is mainly responsible for development of strong interaction forces. High forces will keep T-cells and APCs in tight contact, thereby providing a platform for optimal interaction between TCRs and peptide-MHC complexes.
Division of Molecular Immunology, German Cancer Research Center DKFZ, 69120 Heidelberg, Germany.|Institute of Physiology and Pathophysiology, University of Heidelberg, Germany.
Sabrina
S
Hoffmann
Institute of Biophysical Chemistry, University of Heidelberg, and Dept. of New Materials and Biosystems, Max-Planck-Institute for Metals Research, 70569 Stuttgart, Germany.
Babak H
BH
Hosseini
Institute of Physiology and Pathophysiology, University of Heidelberg, Germany.
Markus
M
Hecker
Institute of Biophysical Chemistry, University of Heidelberg, and Dept. of New Materials and Biosystems, Max-Planck-Institute for Metals Research, 70569 Stuttgart, Germany.
Ilia
I
Louban
Division of Molecular Immunology, German Cancer Research Center DKFZ, 69120 Heidelberg, Germany.
Nadja
N
Bulbuc
Division of Molecular Immunology, German Cancer Research Center DKFZ, 69120 Heidelberg, Germany.
Natalio
N
Garbi
Institute of Immunology, University of Heidelberg, Germany.
Guido H
GH
Wabnitz
Institute of Immunology, University of Heidelberg, Germany.
Yvonne
Y
Samstag
Institute of Biophysical Chemistry, University of Heidelberg, and Dept. of New Materials and Biosystems, Max-Planck-Institute for Metals Research, 70569 Stuttgart, Germany.
Joachim P
JP
Spatz
Division of Molecular Immunology, German Cancer Research Center DKFZ, 69120 Heidelberg, Germany.
Günter J
GJ
Hämmerling
D000938:Antigen-Presenting Cells / Q000276:immunology*; D002460:Cell Line; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D018625:Microscopy, Atomic Force; D009186:Myelin Sheath / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D013053:Spectrophotometry; D013601:T-Lymphocytes / Q000276:immunology*
D010455:Peptides; D018799:Intercellular Adhesion Molecule-1