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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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Accumulation, rolling velocity, and resistance to detachment of SKW3 cells on tenascin. (a) Accumulation of SKW3 cells on  plastic-adsorbed tenascin at various shears. Cells introduced at a concentration of 5 × 106 cells/ml were accumulated for 40 s at the indicated shear stresses in separate experiments. The mean values of two experiments are shown with the range indicated by bars. (b) Rolling velocities of SKW3 cells initially accumulated on plastic-adsorbed tenascin for 40 s at 0.27 dynes/cm2. Velocity measurements were  done on all cells in the field; bars represent SEM for each measurement. (c) Detachment of SKW3 cells bound to plastic-immobilized tenascin under increasing shear. Cells were initially accumulated for 40 s at 0.27 dynes/cm2 and then perfused with cell-free medium for 10 s at  each of the indicated shears. Cell accumulation at the end of the 0.27 dynes/cm2 accumulation step was considered 100% binding; the  numbers of cells remaining at the end of subsequent steps were compared to this value to determine percent detachment. The mean values of two experiments are shown; bars representing the range of measurements are present but are too small to be seen.
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Figure 2: Accumulation, rolling velocity, and resistance to detachment of SKW3 cells on tenascin. (a) Accumulation of SKW3 cells on plastic-adsorbed tenascin at various shears. Cells introduced at a concentration of 5 × 106 cells/ml were accumulated for 40 s at the indicated shear stresses in separate experiments. The mean values of two experiments are shown with the range indicated by bars. (b) Rolling velocities of SKW3 cells initially accumulated on plastic-adsorbed tenascin for 40 s at 0.27 dynes/cm2. Velocity measurements were done on all cells in the field; bars represent SEM for each measurement. (c) Detachment of SKW3 cells bound to plastic-immobilized tenascin under increasing shear. Cells were initially accumulated for 40 s at 0.27 dynes/cm2 and then perfused with cell-free medium for 10 s at each of the indicated shears. Cell accumulation at the end of the 0.27 dynes/cm2 accumulation step was considered 100% binding; the numbers of cells remaining at the end of subsequent steps were compared to this value to determine percent detachment. The mean values of two experiments are shown; bars representing the range of measurements are present but are too small to be seen.

Mentions: The ability of rolling SKW3 cells to accumulate on tenascin decreased with increasing shear stresses (Fig. 2 a) with a profile similar to that seen for SKW3 cells on purified hyaluronan (15). Accumulation was less efficient than was seen with neutrophils (PMN) on E-selectin, where accumulation was still evident at 1.6 dynes/cm2 and declined to zero between 3 to 4.5 dynes/cm2 (48). Rolling velocities of cells on tenascin increased with increasing wall shear stress and were on the order of 10 μm/second (Fig. 2 b), similar to the rolling velocities of PMN on E-selectin (48) but faster than the rolling velocities of SKW3 cells on hyaluronan, which were on the order of 1 μm/s (15). SKW3 cells rolling on tenascin detached with increasing shear stress (Fig. 2 c). SKW3 cells detached from tenascin at lower wall shear stresses than seen with PMN on E-selectin or SKW3 cells on hyaluronan; at 35 dynes/cm2, 96% of adherent SKW3 cells on tenascin had detached, whereas only 60% of PMN on E-selectin and 20% of SKW3 cells on hyaluronan had detached, respectively (15, 48). The ability of cells to accumulate in shear flow, rolling velocity, and resistance to detachment depend not only on intrinsic properties of the receptor–ligand interaction (48) but also to some extent upon the number of binding sites on the substrate, which is a function of the concentration of proteins used during adsorption to plastic. Therefore, the above observations depend on the plating concentrations used in these studies and are not necessarily general properties of the interaction.


Tenascin supports lymphocyte rolling.

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

Accumulation, rolling velocity, and resistance to detachment of SKW3 cells on tenascin. (a) Accumulation of SKW3 cells on  plastic-adsorbed tenascin at various shears. Cells introduced at a concentration of 5 × 106 cells/ml were accumulated for 40 s at the indicated shear stresses in separate experiments. The mean values of two experiments are shown with the range indicated by bars. (b) Rolling velocities of SKW3 cells initially accumulated on plastic-adsorbed tenascin for 40 s at 0.27 dynes/cm2. Velocity measurements were  done on all cells in the field; bars represent SEM for each measurement. (c) Detachment of SKW3 cells bound to plastic-immobilized tenascin under increasing shear. Cells were initially accumulated for 40 s at 0.27 dynes/cm2 and then perfused with cell-free medium for 10 s at  each of the indicated shears. Cell accumulation at the end of the 0.27 dynes/cm2 accumulation step was considered 100% binding; the  numbers of cells remaining at the end of subsequent steps were compared to this value to determine percent detachment. The mean values of two experiments are shown; bars representing the range of measurements are present but are too small to be seen.
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Related In: Results  -  Collection

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Figure 2: Accumulation, rolling velocity, and resistance to detachment of SKW3 cells on tenascin. (a) Accumulation of SKW3 cells on plastic-adsorbed tenascin at various shears. Cells introduced at a concentration of 5 × 106 cells/ml were accumulated for 40 s at the indicated shear stresses in separate experiments. The mean values of two experiments are shown with the range indicated by bars. (b) Rolling velocities of SKW3 cells initially accumulated on plastic-adsorbed tenascin for 40 s at 0.27 dynes/cm2. Velocity measurements were done on all cells in the field; bars represent SEM for each measurement. (c) Detachment of SKW3 cells bound to plastic-immobilized tenascin under increasing shear. Cells were initially accumulated for 40 s at 0.27 dynes/cm2 and then perfused with cell-free medium for 10 s at each of the indicated shears. Cell accumulation at the end of the 0.27 dynes/cm2 accumulation step was considered 100% binding; the numbers of cells remaining at the end of subsequent steps were compared to this value to determine percent detachment. The mean values of two experiments are shown; bars representing the range of measurements are present but are too small to be seen.
Mentions: The ability of rolling SKW3 cells to accumulate on tenascin decreased with increasing shear stresses (Fig. 2 a) with a profile similar to that seen for SKW3 cells on purified hyaluronan (15). Accumulation was less efficient than was seen with neutrophils (PMN) on E-selectin, where accumulation was still evident at 1.6 dynes/cm2 and declined to zero between 3 to 4.5 dynes/cm2 (48). Rolling velocities of cells on tenascin increased with increasing wall shear stress and were on the order of 10 μm/second (Fig. 2 b), similar to the rolling velocities of PMN on E-selectin (48) but faster than the rolling velocities of SKW3 cells on hyaluronan, which were on the order of 1 μm/s (15). SKW3 cells rolling on tenascin detached with increasing shear stress (Fig. 2 c). SKW3 cells detached from tenascin at lower wall shear stresses than seen with PMN on E-selectin or SKW3 cells on hyaluronan; at 35 dynes/cm2, 96% of adherent SKW3 cells on tenascin had detached, whereas only 60% of PMN on E-selectin and 20% of SKW3 cells on hyaluronan had detached, respectively (15, 48). The ability of cells to accumulate in shear flow, rolling velocity, and resistance to detachment depend not only on intrinsic properties of the receptor–ligand interaction (48) but also to some extent upon the number of binding sites on the substrate, which is a function of the concentration of proteins used during adsorption to plastic. Therefore, the above observations depend on the plating concentrations used in these studies and are not necessarily general properties of the interaction.

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