Yoji Shimizu

Lymphocyte interactions with endothelial cells

Adhesion of lymphocytes to endothelium is vital to lymphocyte migration into lymphoid tissue and into inflammatory sites. In this review, Yoji Shimizu and colleagues identify the molecules that mediate lymphocyte-endothelial cell adhesion, describe the underlying principles of lymphocyte migration, and discuss a model of the sequence of events that allow a lymphocyte to successfully attach to endothelium and migrate into the surrounding tissue.

Roles of Adhesion Molecules in T‐Cell Recognition: Fundamental Similarities between Four Integrins on Resting Human T Cells (LFA‐1, VLA‐4, VLA‐5, VLA‐6) in Expression, Binding, and Costimulation

This review summarizes our recent work on expression and function of 4 integrins on resting human CD4+ T cells. Three themes are highlighted: multiplicity of molecular pathways of adhesion, regulation of adhesion, and costimulation by adhesion molecules. Four distinct receptor/ligand interactions have been elucidated: LFA-1/ICAM-1, VLA-5/fibronection, VLA-4/fibronectin, and VLA-6/laminin. Our studies indicate fundamental similarities in function and regulation of these four receptor/ligand interactions: 1) acute activation of the T cell (by CD3/TCR crosslinking or by PMA) induces rapid but transient integrin binding function; and 2) higher expression of each integrin on memory T cells compared to naive T cells results in greater binding of memory cells to each ligand. The identification of T-cell integrins (VLA-4, VLA-5, VLA-6) which interact with ECM components directs attention to the potential importance of T-cell interactions with ECM components which either may be immobilized in ECM or which may act as molecular bridges between cells. The existence of multiple adhesion pathways, of multiple ligands for a single receptor (such as LFA-1/ICAM-1 and LFA-1/ICAM-2), multiple receptors for a single ligand (such as VLA-4/FN and VLA-5/FN), and regulation of ligand expression (ICAM-1) provides opportunities for co-operativity, rebundancy and diversity which the T cell utilizes to exquisitely regulate its adhesive interactions. The thesis that adhesion molecules can be multifunctional receptors that also influence signalling is demonstrated by our findings that each of these integrin receptor/ligand interactions is capable of providing a potent costimulatory signal to CD3-mediated T-cell activation. The importance of interactions of T-cell integrins with their cell surface and ECM ligands is discussed with respect to T-cell migration, differentiation and recognition. Analysis of the precise mechanisms by which T cells regulate and exploit these multiple receptor/ligand interactions and the resulting functional consequences of those interactions will be exciting areas of future research.

Lymphocyte interactions with extracellular matrix.

To mediate an immune response, lymphocytes must be able to interact with and respond to the surrounding extracellular environment. In addition to cell surface molecules that facilitate adhesion of lymphocytes to other cells, recent studies have demonstrated that lymphocytes interact with glycoproteins and glycosaminoglycans that are major components of the extracellular matrix (ECM). Although many receptors mediating the effects of ECM components on lymphocyte function remain poorly defined, a number of lymphocyte ECM receptors have recently been identified; these include members of the integrin family of adhesion molecules as well as structurally unrelated molecules such as CD44 and CD26. Furthermore, as lymphocytes must be able to move between various microenvironments in vivo, they have proved to be an excellent cell type in which to identify and analyze various modes of regulation of cell‐ECM interactions. As with other cell types, the ECM has been shown to have multiple effects on lymphocytes; functional analysis reveals effects of the ECM on lymphocyte migration, recognition/activation, and differentiation. These studies emphasize: 1) the importance of lymphocytes as a model system for identifying and analyzing ECM receptor expression and function, and 2) the multiple roles that the ECM plays in the function of the immune system in vivo.—Shimizu, Y.; Shaw, S. Lymphocyte interactions with extracellular matrix. FASEB J. 5: 2292–2299; 1991.

The WAVE2 Complex Regulates Actin Cytoskeletal Reorganization and CRAC-Mediated Calcium Entry during T Cell Activation

BACKGROUND The engagement of the T cell receptor results in actin cytoskeletal reorganization at the immune synapse (IS) and the triggering of biochemical signaling cascades leading to gene regulation and, ultimately, cellular activation. Recent studies have identified the WAVE family of proteins as critical mediators of Rac1-induced actin reorganization in other cell types. However, whether these proteins participate in actin reorganization at the IS or signaling pathways in T cells has not been investigated. RESULTS By using a combination of biochemical, genetic, and cell biology approaches, we provide evidence that WAVE2 is recruited to the IS, is biochemically modified, and is required for actin reorganization and beta-integrin-mediated adhesion after TCR crosslinking. Moreover, we show that WAVE2 regulates calcium entry at a point distal to PLCgamma1 activation and IP(3)-mediated store release. CONCLUSIONS These data reveal a role for WAVE2 in regulating multiple pathways leading to T cell activation. In particular, this work shows that WAVE2 is a key component of the actin regulatory machinery in T cells and that it also participates in linking intracellular calcium store depletion to calcium release-activated calcium (CRAC) channel activation.

Roles of multiple accessory molecules in T-cell activation

Accessory molecules expressed on T cells can mediate adhesion between T cells and other cells, or the extracellular matrix. The same T-cell accessory molecules participate in a dialogue with their ligands (counter-receptors) on the antigen-presenting cells, and elicit signals that determine the specifics of activation and subsequent differentiation of the T cells and antigen-presenting cells.

Regulation of the PH-domain-containing tyrosine kinase Etk by focal adhesion kinase through the FERM domain

Etk/BMX, a member of the Btk family of tyrosine kinases, is highly expressed in cells with great migratory potential, including endothelial cells and metastatic carcinoma cell lines. Here, we present evidence that Etk is involved in integrin signalling and promotes cell migration. The activation of Etk by extracellular matrix proteins is regulated by FAK through an interaction between the PH domain of Etk and the FERM domain of FAK. The lack of Etk activation by extracellular matrix in FAK-null cells could be restored by co-transfection with wild-type FAK. Disrupting the interaction between Etk and FAK diminished the cell migration promoted by either kinase. Furthermore, inhibiting Etk expression in metastatic carcinoma cell lines with an antisense oligonucleotide blocks integrin-mediated migration of these cells. Taken together, our data indicate the essential role of the interaction of the PH domain of Etk and the FERM domain of FAK in integrin signalling.

Integrins in the Immune System

Publisher Summary This chapter discusses the principles governing the integrins, their capacity to recognize ligands, cellular regulation of integrin function, and aspects of integrin function in the immune system. The chapter brings these general principles into focus to understand the immune system and to describe how perturbation of these receptors can be used to modulate immunological responses. Integrins are heterodimers formed by combination of 17 α and 8 β subunits in humans and are important because they play key roles in the cell migration, cell adhesion, control of differentiation, and critical decisions specifying cell proliferation and programmed cell death. The remarkable information storage and retrieval capacities of the immune system employ the general machinery involved in vertebrate development. Thus, integrins play crucial roles in modulating signals generated by clonotypic receptors and as effectors in processes such as cytolytic killing and phagocytosis.

β-Actin specifically controls cell growth, migration, and the G-actin pool

Targeted deletion of Actb demonstrates that the β-actin gene, in contrast to the γ-actin gene, is an essential gene uniquely required for cell growth and migration. Cell motility and growth defects in β-actin–knockout primary cells are due to a specific role for β-actin in regulating gene expression through control of the cellular G-actin pool.

A novel function for the Tec family tyrosine kinase Itk in activation of β1 integrins by the T-cell receptor

Stimulation of T cells via the CD3–T‐cell receptor (TCR) complex results in rapid increases in β1 integrin‐mediated adhesion via poorly defined intracellular signaling events. We demonstrate that TCR‐mediated activation of β1 integrins requires activation of the Tec family tyrosine kinase Itk and phosphatidylinositol 3‐kinase (PI 3‐K)‐dependent recruitment of Itk to detergent‐insoluble glycosphingolipid‐enriched microdomains (DIGs) via binding of the pleckstrin homology domain of Itk to the PI 3‐K product PI(3,4,5)‐P3. Activation of PI 3‐K and the src family kinase Lck, via stimulation of the CD4 co‐receptor, can initiate β1 integrin activation that is dependent on Itk function. Targeting of Itk specifically to DIGs, coupled with CD4 stimulation, can also activate β1 integrin function independently of TCR stimulation. Changes in β1 integrin function mediated by TCR activation of Itk are also accompanied by Itk‐dependent modulation of the actin cytoskeleton. Thus, TCR‐mediated activation of β1 integrins involves membrane relocalization and activation of Itk via coordinate action of PI 3‐K and a src family tyrosine kinase.

Cutting Edge: T Cell Migration Regulated by CXCR4 Chemokine Receptor Signaling to ZAP-70 Tyrosine Kinase

Chemokines regulate the homeostatic trafficking of lymphocytes and lymphocyte influx into sites of injury and inflammation. The signaling pathways by which chemokine receptors regulate lymphocyte migration remain incompletely characterized. We demonstrate that Jurkat T cells lacking the ZAP-70 tyrosine kinase exhibit reduced migration in response to the CXCR4 ligand CXCL12 when compared with wild-type Jurkat T cells. Expression of wild-type, but not kinase-inactive, ZAP-70 resulted in enhanced migration of ZAP-70-deficient Jurkat T cells. The tyrosine residue at position 292 in the interdomain B region of ZAP-70 exerts a negative regulatory effect on ZAP-70-dependent migration. Stimulation of Jurkat T cells with CXCL12 also resulted in ZAP-70-dependent tyrosine phosphorylation of the Src homology 2 domain-containing leukocyte protein of 76 kDa (SLP-76) adapter protein. Although CXCL12-dependent migration of SLP-76-deficient Jurkat T cells was impaired, re-expression of SLP-76 did not enhance migration. These results suggest a novel function for ZAP-70, but not SLP-76, in CXCR4 chemokine receptor signaling in human T cells.