Contact formation of T cells with antigen presenting cells results in the engagement of T cell receptors (TCRs), recruitment and aggregation of signaling proteins into microclusters and ultimately, T cell activation. During this process, T cells undergo dramatic changes in cell shape and reorganization of the cytoskeleton. While the importance of the cytoskeleton in T cell activation is well known, the dynamics of the actin cytoskeleton and how it correlates with signaling clusters during the early stages of spreading is not well understood. Here, we used total internal reflection fluorescence microscopy to study the dynamics of actin reorganization during Jurkat T cell spreading and the role of integrin ligation by the adhesion molecule, vascular cell adhesion molecule (VCAM), in modulating actin dynamics. We found that when T cells spread on anti-CD3 antibody-coated glass surfaces, the cell edge exhibited repeated protrusions and retractions, which were driven by wave like patterns of actin that emerged from signaling microclusters. Addition of VCAM on the activating substrate altered the dynamics of actin both globally and locally, leading to a smooth expansion of the cell edge and the disappearance of waves. Our results suggest that the actin cytoskeleton in Jurkat cells is capable of organizing into spatial patterns initiated by TCR signaling and regulated by integrin signaling.
Keywords: actin waves; integrins; lymphocytes; total internal reflection fluorescence.
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