Determination of Junction Avidity of Cytolytic T Cell and Target Cell

Sung, Kuo‐Li Paul; Sung, Lanping Amy; Crimmins, Mary A. V.; Burakoff, Steven J.; Chien, Shu

doi:10.1126/science.3491426

Cited by 119 publications

(90 citation statements)

References 23 publications

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“…The experimental technique is based on a modification of a dual-micropipette assay (54), which permits manipulation of individual cells to form controlled contacts. Briefly, the experiments were performed on the stage of a Leica inverted epifluorescence microscope positioned on an anti-vibration platform and equipped with a cooled charge-coupled device Hamamatsu C5985 or Nikon Coolpix 5000 camera, a digitally controlled heating stage and 10ϫ and 63ϫ objectives.…”

Section: Methodsmentioning

confidence: 99%

Separation Force Measurements Reveal Different Types of Modulation of E-cadherin-based Adhesion by Nectin-1 and -3

Martinez-Rico

Pincet

Perez

et al. 2005

Journal of Biological Chemistry

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Nectins are Ca2؉ -independent cell adhesion molecules found at cadherin-based adherens junctions. We used a dual pipette assay that measures the forces required to separate cell doublets to determine how nectins affect the formation and strength of cell-cell adhesion. Less force was required to separate doublets of L cells expressing nectin-1 or nectin-3 than to separate doublets of E-cadherin-expressing cells. Heterodimers formed between cells expressing nectin-1 or nectin-3 adhered more strongly than homodimers. Nectin-3 that does not trans-interact with nectin-1 inhibited E-cadherin-mediated adhesion. However, the extracellular fragment of nectin-1 did not have an agonistic effect on E-cadherindependent cell adhesion when it trans-interacted with nectin-3, expressed at high levels in cells. In contrast, the extracellular fragment of nectin-3 had a significant agonistic effect on cadherin-based adhesion when it interacted with endogenous nectin-1, expressed at low levels in cells. Our results indicate that E-cadherin is the key molecule involved in cell adhesion and that the regulation of E-cadherin-based adhesion involving cellular nectin-1 trans-interacting with nectin-3 is qualitatively different from that involving cellular nectin-3 trans-interacting with nectin-1 and depends on the nectin levels expressed by cells.Cadherin-based cell adhesion is a critical determinant of tissue architecture in developing and adult metazoan organisms (1-3). Cadherins (4 -10) are single-pass, integral membrane proteins of 115-130 kDa. They belong to the family of Ca 2ϩ -dependent cell adhesion molecules, which affect cell morphology, architecture, and function (3, 11-13). More than 80 cadherins have been identified to date, each of which is expressed in a wide variety of cells, including epithelial and non-epithelial cells (3, 13). Classical cadherins function as multiprotein complexes: the ectodomains mediate homophilic adhesive binding (2,14), whereas the intracellular C-terminal domains associate with catenins at the site of adherens junctions (AJs), 1 forming cadherin-catenin complexes (15, 16). The ␤-and ␥-catenins interact with the cytoplasmic tail of E-cadherin, forming a complex that is coupled to the actin cytoskeleton through ␣-catenin (also an actin-binding protein (8, 17)) via its interactions with a number of actin-binding proteins, such as ␣-actinin and vinculin, or by directly binding to actin itself (18 -21). These complexes are the functional cell adhesive units (22). The interaction between cadherin and catenin also requires Ca 2ϩ (22)(23)(24)(25)(26) and is regulated by GTPases (17, 27). The association of E-cadherin with the actin cytoskeleton through these peripheral membrane proteins strengthens the cell-cell adhesion activity of E-cadherin (2, 18).Nectins belong to a new family of integral membrane proteins found in most AJs (28). Nectins share structural and sequence features and are characterized by three Ig-like domains in their extracellular portion. Nectins are widely expressed in tissues and cel...

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Section: Methodsmentioning

confidence: 99%

Separation Force Measurements Reveal Different Types of Modulation of E-cadherin-based Adhesion by Nectin-1 and -3

Martinez-Rico

Pincet

Perez

et al. 2005

Journal of Biological Chemistry

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“…To confirm and quantify this enhanced adhesion with the CX3CR1-IM variant, we used another cell-cell adhesion assay, the dual pipette aspiration technique, previously used to verify CTL target adhesion (32). Briefly, this method consists in determining the force required to dissociate a pair formed by two cells brought into contact by micropipettes.…”

Section: Adhesive Capacities Of the Cx3cr1 Variantsmentioning

confidence: 99%

“…Cells were manipulated with two micropipettes, each held in its own micromanipulator and connected to a combined hydraulic/pneumatic system that provided the necessary control of the aspiration force applied to the cells. The protocol we used is very similar to that of Chien and co-workers (32). Two cells, collected by gentle aspiration onto the tip of each pipette (cell number 1 in pipette A, cell 2 in pipette B), were brought into contact through the use of the micromanipulators and allowed to remain in contact for different periods of time (Fig.…”

mentioning

confidence: 99%

Enhanced Adhesive Capacities of the Naturally Occurring Ile249–Met280 Variant of the Chemokine Receptor CX3CR1

Daoudi

Lavergne

Garin

et al. 2004

Journal of Biological Chemistry

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. We report here that peripheral blood mononuclear cells (PBMC) from individuals with the CX3CR1-IM haplotype adhered more potently to membrane-bound CX3CL1 than did PBMC from homozygous CX3CR1-VT donors. Similar excess adhesion was observed with CX3CR1-IMtransfected human embryonic kidney (HEK) cell lines tested with two different methods: the parallel plate laminar flow chamber and the dual pipette aspiration technique. Suppression of the extra adhesion in the presence of pertussis toxin indicates that G-protein mediated the underlying transduction pathway, in contrast to the G-protein-independent adhesion previously described for CX3CR1-VT. Surprisingly, HEK and PBMC that expressed CX3CR1-IM and -VT were indistinguishable when tested with the soluble form of CX3CL1 for chemotaxis, calcium release, and binding capacity. In conclusion, only the membrane-anchored form of CX3CL1 functionally discriminated between these two allelic isoforms of CX3CR1. These results suggest that each form of this ligand may lead to a different signaling pathway. The extra adhesion of CX3CR1-IM may be related to immune defenses and to atherogenesis, both of which depend substantially on adhesive intercellular events.Adhesion, a critical stage in cell trafficking and migration (1, 2), requires the presence of numerous adhesion molecules, such as integrins, that need divalent ions to function. Recent studies show that two chemokines, namely, CX3CL1 and CXCL16, are not only chemoattractant as soluble molecules, but also function as adhesion molecules since they are membrane-anchored, regardless of the presence of divalent ions (3-7). The best known of these is CX3CL1, also called fractalkine. It is expressed on the surface of many types of cells, in particular interleukin-1-and tumor necrosis factor-activated endothelial (4) and dendritic cells (8), as a membrane molecule containing the classic chemokine domain, a mucin-like stalk, and a transmembrane domain tethering it to the cell membrane. CX3CL1 may be cleaved by TACE and released from cells (9, 10). In this soluble form, it behaves like a chemoattractant molecule, just as other chemokines do. The transmembrane feature of the native CX3CL1 protein, combining it with its receptor, CX3CR1, produces a strongly adhesive pair (4, 5, 11) that mediates the rapid capture and firm adhesion of leukocytes. Because this activity persists in the absence of divalent cations, it is thought to be independent of integrins (5,11,12). This adhesive feature is also independent of the G i pathway, since it is still present after pertussis toxin (PTX) 1 treatment (4, 5, 11). The CX3CR1 molecule is expressed on leukocytes, especially monocytes (4) and cytotoxic cells (13,14), on dendritic cells (15), and on neurons and microglial cells (16,17). Recently, we identified two common polymorphisms in strong linkage disequilibrium in the CX3CR1 gene: V249I and T280M (18). We also found that these mutations are associated with more rapid progression to AIDS (18,19), although two studies have failed to confirm t...

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“…The step-pressure technique, which was developed by Sung et al in 1986, is the first micropipettebased technique that was employed for quantifying celladhesion strength (17). The major components of a micropipette-manipulation system are micropipettes (cylindrical glass tubes with internal diameters of a few micrometers), micromanipulators (for controlling the positions of the micropipettes in an experimental chamber), and manometers (for controlling the pressures within the micropipettes).…”

Section: The Step-pressure Technique (Spt)mentioning

confidence: 99%

“…Ten years after this pioneering effort, Rand and Burton further refined the elastimeter and applied it to the study of the viscoelastic properties of the erythrocyte membrane (14,15). In the 1970s and 80s, the micropipettemanipulation system was significantly improved at several laboratories and applied to a myriad of studies on the mechanics of erythrocytes, leukocytes, endothelial cells, and lipid vesicles or liposomes (16)(17)(18)(19)(20). In these early studies, either a point force or a suction pressure over a small area was imposed on a single cell or liposome surface.…”

Section: Introductionmentioning

confidence: 99%

Quantifying cell-adhesion strength with micropipette manipulation: principle and application

Shao¹

2004

Front Biosci

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Quantifying cell-adhesion strength is of great importance in biology and medicine. Cell-adhesion strength can be characterized by separating two adherent cells and determining the force required to do so, or by measuring the lifetime of a receptor-ligand bond that mediates cell adhesion. To this end, several micropipette-based experimental techniques that operate at both cellular and molecular levels have been developed over the past few decades. In this review, we provide an overview of three of these techniques, i.e., the step-pressure technique (SPT), the biomembraneforce probe (BFP), and the micropipette-aspiration technique (MAT). More detailed discussion will be given about the requirements and applications of the MAT.

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Determination of Junction Avidity of Cytolytic T Cell and Target Cell

Cited by 119 publications

References 23 publications

Separation Force Measurements Reveal Different Types of Modulation of E-cadherin-based Adhesion by Nectin-1 and -3

Separation Force Measurements Reveal Different Types of Modulation of E-cadherin-based Adhesion by Nectin-1 and -3

Enhanced Adhesive Capacities of the Naturally Occurring Ile249–Met280 Variant of the Chemokine Receptor CX3CR1

Quantifying cell-adhesion strength with micropipette manipulation: principle and application

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