Limits...
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.

Show MeSH

Related in: MedlinePlus

Rolling on tenascin is smoother than rolling on E-selectin at all shear stresses examined. (a–c) Displacement and velocity profiles of cells rolling on tenascin and E-selectin at 2.7 dynes/cm2 shear stress. (c–e) Change in velocity profiles of cells rolling on tenascin  and E-selectin at 10 μm/s. The variance of velocity measurements is as shown.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2139881&req=5

Figure 7: Rolling on tenascin is smoother than rolling on E-selectin at all shear stresses examined. (a–c) Displacement and velocity profiles of cells rolling on tenascin and E-selectin at 2.7 dynes/cm2 shear stress. (c–e) Change in velocity profiles of cells rolling on tenascin and E-selectin at 10 μm/s. The variance of velocity measurements is as shown.

Mentions: KG1a cells on tenascin at 290 sites/μm2 rolled at similar velocities to KG1a cells on a plating dilution of 1:500 E-selectin (Fig. 6 b). During these experiments it was noted that KG1a cells rolling on E-selectin appeared to move in a series of jerks, as previously noted for rolling on selectins (3), whereas cells attached to tenascin rolled more smoothly. We quantitated variation in rolling velocity using separate conditions under which either the wall shear stress or the average rolling velocity were identical on the three substrates. The position of representative rolling cells was measured every 0.033 s. For each condition, both the displacement (Fig. 7, top graph) and the velocity (Fig. 7, bottom graph) are shown. A relatively straight tracing on the displacement versus time graph indicates a cell that rolled smoothly at a constant velocity, while a tracing with changes in slope indicates a cell that rolled in a more jerky manner. Similarly, the range of velocities for a smoothly rolling cell is small, while the intermittent motion of a jerky cell is reflected in a larger range of rolling velocities. This was quantified by calculating the variance in cell velocities for cells rolling on the three substrates. Cells rolling on tenascin had a variance in velocities of 40.4, whereas cells rolling on 1:1,000 and 1:500 dilutions of E-selectin had larger variances, 425 and 104, respectively. Therefore, cells rolled more smoothly on tenascin at 2.7 dynes/cm2 shear stress than cells rolling on either site density of E-selectin (Fig. 7, a–c). To compare cells rolling at a similar velocity of 10 μm/s, cells rolling at 2.7 dynes/cm2 shear stress for E-selectin (1:1,000) and at 10 dynes/cm2 for E-selectin (1: 500) and tenascin were compared (Figs. 6 b and 7, d–f). Cells bound to tenascin rolled more smoothly than cells bound to E-selectin when rolling at similar velocities. The variance in cell velocities was 75.8 for cells rolling on tenascin and 425 and 246 for cells rolling on 1:1,000 and 1:500 dilutions of E-selectin, respectively. These direct comparisons suggest that the smoother rolling observed with tenascin is an inherent characteristic of the molecular interaction of tenascin with its cellular receptor.


Tenascin supports lymphocyte rolling.

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

Rolling on tenascin is smoother than rolling on E-selectin at all shear stresses examined. (a–c) Displacement and velocity profiles of cells rolling on tenascin and E-selectin at 2.7 dynes/cm2 shear stress. (c–e) Change in velocity profiles of cells rolling on tenascin  and E-selectin at 10 μm/s. The variance of velocity measurements is as shown.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 7: Rolling on tenascin is smoother than rolling on E-selectin at all shear stresses examined. (a–c) Displacement and velocity profiles of cells rolling on tenascin and E-selectin at 2.7 dynes/cm2 shear stress. (c–e) Change in velocity profiles of cells rolling on tenascin and E-selectin at 10 μm/s. The variance of velocity measurements is as shown.
Mentions: KG1a cells on tenascin at 290 sites/μm2 rolled at similar velocities to KG1a cells on a plating dilution of 1:500 E-selectin (Fig. 6 b). During these experiments it was noted that KG1a cells rolling on E-selectin appeared to move in a series of jerks, as previously noted for rolling on selectins (3), whereas cells attached to tenascin rolled more smoothly. We quantitated variation in rolling velocity using separate conditions under which either the wall shear stress or the average rolling velocity were identical on the three substrates. The position of representative rolling cells was measured every 0.033 s. For each condition, both the displacement (Fig. 7, top graph) and the velocity (Fig. 7, bottom graph) are shown. A relatively straight tracing on the displacement versus time graph indicates a cell that rolled smoothly at a constant velocity, while a tracing with changes in slope indicates a cell that rolled in a more jerky manner. Similarly, the range of velocities for a smoothly rolling cell is small, while the intermittent motion of a jerky cell is reflected in a larger range of rolling velocities. This was quantified by calculating the variance in cell velocities for cells rolling on the three substrates. Cells rolling on tenascin had a variance in velocities of 40.4, whereas cells rolling on 1:1,000 and 1:500 dilutions of E-selectin had larger variances, 425 and 104, respectively. Therefore, cells rolled more smoothly on tenascin at 2.7 dynes/cm2 shear stress than cells rolling on either site density of E-selectin (Fig. 7, a–c). To compare cells rolling at a similar velocity of 10 μm/s, cells rolling at 2.7 dynes/cm2 shear stress for E-selectin (1:1,000) and at 10 dynes/cm2 for E-selectin (1: 500) and tenascin were compared (Figs. 6 b and 7, d–f). Cells bound to tenascin rolled more smoothly than cells bound to E-selectin when rolling at similar velocities. The variance in cell velocities was 75.8 for cells rolling on tenascin and 425 and 246 for cells rolling on 1:1,000 and 1:500 dilutions of E-selectin, respectively. These direct comparisons suggest that the smoother rolling observed with tenascin is an inherent characteristic of the molecular interaction of tenascin with its cellular receptor.

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