Zhengyu Ma

Mechanical Force in T Cell Receptor Signal Initiation

T cell activation is the result of direct cell-cell contact between T cells and antigen presenting cells (APCs), and of interactions between membrane-bound ligands and receptors at the contact interface, the “immunological synapse.” Model APCs based upon supported fluid lipid bilayers have been used to dissect these complex molecular interactions and to facilitate real-time microscopic observations. Nearly all studies have used liposome fusion-based methods to make supported bilayers, and the biophysical properties of these membranes were not characterized in detail. Here, using both Langmuir-Blodgett and liposome fusion techniques, we explored five different methods of lipid bilayer preparation on glass, mica, or dextran cushion substrates and characterized the stability, homogeneity, and fluidity of the bilayers with fluorescence microscopy and fluorescence recovery after photobleaching (FRAP). Most combinations of techniques and substrates led to unsatisfactory results, notably, a lack of homogeneity for liposome fusion on glass, low stability of bilayers on mica, and loss of fluidity of dextran-cushioned bilayers in solutions containing protein. To overcome these deficits, we developed a technique that combines liposome fusion on glass and thermally enhanced bilayer expansion. The newly expanded pristine bilayer showed high degrees of stability, homogeneity, and fluidity. MHC and ICAM-1 molecules anchored on the bilayer diffused freely and stimulated T cell calcium flux and adhesion, respectively. Microsc. Res. Tech., 2011.

At the interface between T cell and antigen-presenting cell (APC), peptide antigen presented by MHC (pMHC) binds to the T cell receptor (TCR) and initiates signaling. The mechanism of TCR signal initiation, or triggering, remains unclear. An interesting aspect of this puzzle is that although soluble agonist pMHCs cannot trigger TCR even at high concentrations, the same ligands trigger TCR very efficiently on the surface of APCs. Here, using lipid bilayers or plastic-based artificial APCs with defined components, we identify the critical APC-associated factors that confer agonist pMHCs with such potency. We found that CD4⁺ T cells are triggered by very low numbers of monomeric agonist pMHCs anchored on fluid lipid bilayers or fixed plastic surfaces, in the absence of any other APC surface molecules. Importantly, on bilayers, plastic surfaces, or real APCs, endogenous pMHCs did not enhance TCR triggering. TCR triggering, however, critically depended upon the adhesiveness of the surface and an intact T cell actin cytoskeleton. Based on these observations, we propose the receptor deformation model of TCR triggering to explain the remarkable sensitivity and specificity of TCR triggering. Using the T cell receptor (TCR) as a sensor, T cells of the immune system constantly migrate in lymphoid organs and probe the surface of antigen-presenting cells (APCs) for foreign antigens, a sign of pathogen infection. Antigen binding by TCRs leads to T cell activation and subsequent immune response to combat the pathogens. Interestingly, although T cells respond well to antigens on APCs, they do not recognize the same antigens in solution. What is it that makes antigens on APCs recognizable? To address this, we used lipid bilayers and plastic surfaces to construct artificial APCs with defined antigen number, composition, and configuration. We found that T cells respond to very few individual foreign antigens on artificial APCs, and contrary to some current opinion, formation of antigen clusters on APCs is not required for antigen recognition by T cells. TCR triggering, however, requires T cell adhesion to the APC surface and then occurs only if the T cells are able to move. We propose that at the dynamic T cell–APC interface, antigen on APCs activates T cells by applying force to the TCR and deforming its structure, which cannot be achieved by soluble antigens due to their lack of anchorage.

Through T cell receptors (TCRs), T cells can detect and respond to very small numbers of foreign peptides among a huge number of self‐peptides presented by major histocompatibility complexes (pMHCs) on the surface of antigen‐presenting cells (APCs). How T cells achieve such remarkable sensitivity and specificity through pMHC‐TCR binding is an intensively pursued issue in immunology today;the key question is how pMHC‐TCR binding initiates, or triggers, a signal from TCRs. Multiple competing models have been proposed, none of which fully explains the sensitivity and specificity of TCR triggering. What has been omitted from existing theories is that the pMHC‐TCR interaction at the T cell/APC interface must be under constant mechanical stress, due to the dynamic nature of cell‐cell interaction. Taking this condition into consideration, we propose the receptor deformation model of TCR triggering. In this model, TCR signaling is initiated by conformational changes of the TCR/CD3 complex, induced by a pulling force originating from the cytoskeleton and transmitted through pMHC‐TCR binding interactions with enough strength to resist rupture. By introducing mechanical force into a model of T cell signal initiation, the receptor deformation model provides potential mechanistic solutions to the sensitivity and specificity of TCR triggering. Ma, Z., Janmey, P. A., Finkel, T. H. The receptor deformation model of TCR triggering. FASEB J. 22, 1002–1008 (2008)