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Tenascin supports lymphocyte rolling.

Clark RA, Erickson HP, Springer TA - J. Cell Biol. (1997)

Bottom Line: Tenascin has been reported to have both adhesive and anti-adhesive effects in static assays.When compared to rolling of the same cell type on E-selectin, rolling on tenascin was found to be smoother at all shear stresses tested, suggesting that cells formed a larger number of bonds on the tenascin substrate than on the E-selectin substrate.When protein plating densities were adjusted to give similar profiles of cell detachment under increasing shears, the density of tenascin was 8.5-fold greater than that of E-selectin.

View Article: PubMed Central - PubMed

Affiliation: The Center for Blood Research and Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
Tenascin is a large extracellular matrix molecule expressed at specific sites in the adult, including immune system tissues such as the bone marrow, thymus, spleen, and T cell areas of lymph nodes. Tenascin has been reported to have both adhesive and anti-adhesive effects in static assays. We report here that tenascin supports the tethering and rolling of lymphocytes and lymphoblastic cell lines under flow conditions. Binding was calcium dependent and was not inhibited by treatment of lymphocytes with O-glycoprotease or a panel of glycosidases including neuraminidase and heparitinase but was inhibited by treatment of cells with proteinase K. Binding was to the fibrinogen-like terminal domain of tenascin as determined by antibody blocking studies and binding to recombinant tenascin proteins. When compared to rolling of the same cell type on E-selectin, rolling on tenascin was found to be smoother at all shear stresses tested, suggesting that cells formed a larger number of bonds on the tenascin substrate than on the E-selectin substrate. When protein plating densities were adjusted to give similar profiles of cell detachment under increasing shears, the density of tenascin was 8.5-fold greater than that of E-selectin. Binding to tenascin was not dependent on any molecules previously identified as tenascin receptors and is likely to involve a novel tenascin receptor on lymphocytes. We postulate that the ability of tenascin to support lymphocyte rolling may reflect its ability to support cell migration and that this interaction may be used by lymphocytes migrating through secondary lymphoid organs.

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Ability of recombinant tenascin proteins to support  SKW3 cell rolling. Cells were accumulated for 40 s at 0.27 dynes/ cm2 shear stress; adhesion events were counted throughout this  40 s period. “Accumulation” represents cells that rolled a distance of at least four cell diameters. “Tethering events” were defined as cells that tethered under flow and remained bound to the  substrate for at least one second. Mean values of two experiments  are shown, and the range is indicated by bars.
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Figure 5: Ability of recombinant tenascin proteins to support SKW3 cell rolling. Cells were accumulated for 40 s at 0.27 dynes/ cm2 shear stress; adhesion events were counted throughout this 40 s period. “Accumulation” represents cells that rolled a distance of at least four cell diameters. “Tethering events” were defined as cells that tethered under flow and remained bound to the substrate for at least one second. Mean values of two experiments are shown, and the range is indicated by bars.

Mentions: To determine if recombinant tenascin proteins could support rolling, we immobilized the proteins on plastic precoated with nonspecific IgG and HSA (as described in Materials and Methods) and used them as substrates in the rolling assay. Rolling of SKW3 cells was observed on TNfbg and TNfn6-8fbg but not on the other recombinant fragments (Fig. 5). Overall levels of accumulation on recombinant TNfbg was ∼10% of that observed with the intact protein and was therefore considerably less efficient. Binding to TNfn6-8fbg was even less efficient, perhaps reflecting reduced binding to IgG-coated plates. Transient tethering, defined as cells that bound under flow and remained adherent to the substrate for at least 1 s, was observed at low levels for TNfnA-D, TNfn1-5, and TNfn6-8 and not for control substrates. Transient tethering of cells to TNfn6-8fbg and TNfbg was two- and fivefold higher, respectively, than that seen with the other proteins. The recombinant tenascin fragments expressed in bacteria are not glycosylated, and thus carbohydrate on tenascin is not required for rolling adhesion. The observation that recombinant proteins containing the fibrinogen-like domain of tenascin can support cell rolling, and the ability of antibodies to this region to block rolling suggests that this domain supports rolling of SKW3 cells.


Tenascin supports lymphocyte rolling.

Clark RA, Erickson HP, Springer TA - J. Cell Biol. (1997)

Ability of recombinant tenascin proteins to support  SKW3 cell rolling. Cells were accumulated for 40 s at 0.27 dynes/ cm2 shear stress; adhesion events were counted throughout this  40 s period. “Accumulation” represents cells that rolled a distance of at least four cell diameters. “Tethering events” were defined as cells that tethered under flow and remained bound to the  substrate for at least one second. Mean values of two experiments  are shown, and the range is indicated by bars.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2139881&req=5

Figure 5: Ability of recombinant tenascin proteins to support SKW3 cell rolling. Cells were accumulated for 40 s at 0.27 dynes/ cm2 shear stress; adhesion events were counted throughout this 40 s period. “Accumulation” represents cells that rolled a distance of at least four cell diameters. “Tethering events” were defined as cells that tethered under flow and remained bound to the substrate for at least one second. Mean values of two experiments are shown, and the range is indicated by bars.
Mentions: To determine if recombinant tenascin proteins could support rolling, we immobilized the proteins on plastic precoated with nonspecific IgG and HSA (as described in Materials and Methods) and used them as substrates in the rolling assay. Rolling of SKW3 cells was observed on TNfbg and TNfn6-8fbg but not on the other recombinant fragments (Fig. 5). Overall levels of accumulation on recombinant TNfbg was ∼10% of that observed with the intact protein and was therefore considerably less efficient. Binding to TNfn6-8fbg was even less efficient, perhaps reflecting reduced binding to IgG-coated plates. Transient tethering, defined as cells that bound under flow and remained adherent to the substrate for at least 1 s, was observed at low levels for TNfnA-D, TNfn1-5, and TNfn6-8 and not for control substrates. Transient tethering of cells to TNfn6-8fbg and TNfbg was two- and fivefold higher, respectively, than that seen with the other proteins. The recombinant tenascin fragments expressed in bacteria are not glycosylated, and thus carbohydrate on tenascin is not required for rolling adhesion. The observation that recombinant proteins containing the fibrinogen-like domain of tenascin can support cell rolling, and the ability of antibodies to this region to block rolling suggests that this domain supports rolling of SKW3 cells.

Bottom Line: Tenascin has been reported to have both adhesive and anti-adhesive effects in static assays.When compared to rolling of the same cell type on E-selectin, rolling on tenascin was found to be smoother at all shear stresses tested, suggesting that cells formed a larger number of bonds on the tenascin substrate than on the E-selectin substrate.When protein plating densities were adjusted to give similar profiles of cell detachment under increasing shears, the density of tenascin was 8.5-fold greater than that of E-selectin.

View Article: PubMed Central - PubMed

Affiliation: The Center for Blood Research and Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
Tenascin is a large extracellular matrix molecule expressed at specific sites in the adult, including immune system tissues such as the bone marrow, thymus, spleen, and T cell areas of lymph nodes. Tenascin has been reported to have both adhesive and anti-adhesive effects in static assays. We report here that tenascin supports the tethering and rolling of lymphocytes and lymphoblastic cell lines under flow conditions. Binding was calcium dependent and was not inhibited by treatment of lymphocytes with O-glycoprotease or a panel of glycosidases including neuraminidase and heparitinase but was inhibited by treatment of cells with proteinase K. Binding was to the fibrinogen-like terminal domain of tenascin as determined by antibody blocking studies and binding to recombinant tenascin proteins. When compared to rolling of the same cell type on E-selectin, rolling on tenascin was found to be smoother at all shear stresses tested, suggesting that cells formed a larger number of bonds on the tenascin substrate than on the E-selectin substrate. When protein plating densities were adjusted to give similar profiles of cell detachment under increasing shears, the density of tenascin was 8.5-fold greater than that of E-selectin. Binding to tenascin was not dependent on any molecules previously identified as tenascin receptors and is likely to involve a novel tenascin receptor on lymphocytes. We postulate that the ability of tenascin to support lymphocyte rolling may reflect its ability to support cell migration and that this interaction may be used by lymphocytes migrating through secondary lymphoid organs.

Show MeSH
Related in: MedlinePlus