Probing T cell membrane organization using dimeric MHC-Ig complexes

In this report, we review a novel method for probing the membrane organization of T cells using dimeric major histocompatibility complexes (MHC), MHC-Ig. MHC-Ig complexes are useful reagents for quantitative analysis of binding data since their valency is controlled. These complexes can be easily labeled and loaded with a variety of peptides. A binding assay using these dimers and quantitative analysis of the MHC-Ig dimer-T cell binding curves is described in detail. Using this approach, we show that the organization of TCR on activated T cells is different from TCR organization on nai;ve T cells. The implications of these findings are discussed with regards to current models of T cell recognition. This analysis offers insight into how T cell controls their biological range of responsiveness. Specifically, these findings reveal the biophysical basis of the ability of activated T cells to recognize low amounts of antigen independent of costimulation.