TCR-Mediated Adhesion of T Cell Hybridomas to Planar Bilayers Containing Purified MHC Class II/Peptide Complexes and Receptor Shedding during Detachment

Michael L. Dustin, James M. Miller, Sheila Ranganath, Dario A.A. Vignali, Nicholas J. Viner, Christopher A. Nelson, Emil R. Unanue

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41 Scopus citations

Abstract

T cell recognition of foreign Ag/MHC class Il complexes is sensitive down to ∼100 complexes per cell or ∼0.2 complexes/μm2. To better understand the physical basis of the recognition stage of Ag presentation, we examined adhesion of the lysozymespecific T cell hybridoma, 3A9, to artificial bilayers containing covalent MHC class II/peptide complexes or adhesion molecules. Adhesion of 3A9 cells required a superphysiologic density of the MHC class II/peptide complex and was partly dependent on CD4; cells adhered but did not crawl. No adhesion was observed to bilayers containing MHC class II molecules without the lysozyme peptide. Activated 3A9 cells adhered and crawled on bilayers containing ICAM-1. The physical strength of contacts was tested with fluid shear. 3A9 cells adherent to bilayers containing MHC class II/peptide complexes shed their contact, which remained on the substrate and contained TCR. In contrast, 3A9 cells peeled from the ICAM-1 bilayer, and held firmly on LFA-1 bilayers in a manner dependent on filamentous actin. When ICAM-1 and the MHC/peptide complexes were combined, the 3A9 cells adhered tightly and spread, but did not crawl, on the bilayers and TCR clustered at the center of the contact area. Physiologically, the TCR is unlikely to directly initiate adhesion. TCR clusters formed with the assistance of adhesion mechanisms may have to be shed to allow de-adhesion, and this may contribute to TCR down-regulation.

Original languageEnglish
Pages (from-to)2014-2021
Number of pages8
JournalJournal of Immunology
Volume157
Issue number5
StatePublished - Sep 1 1996

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