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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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Cell accumulation on plastic-adsorbed tenascin. (a) Ability of various cell types to accumulate on tenascin under flow. Cells  were accumulated for 40 s each at 0.27 and 0.53 dynes/cm2. Cells were counted after the 0.53 dynes/cm2 accumulation step; all adherent  cells were rolled. One of two representative experiments is shown with the range indicated by bars. (b) Effects of inhibitors and enzyme  treatments on SKW3 cell accumulation on tenascin. SKW3 cells were treated for 10 min at 25°C with either 0.1% azide and 50 mM  2-deoxyglucose (Azide+DOG), inclusion of 5 mM EDTA or 10 mM EGTA and 1 mM Mg2+ in the assay, or treatment for 40 min at 37°  with chondroitinase ABC, heparitinase, hyaluronidase, neuraminidase, neuraminidase and exo-β-galactosidase, O-glycoprotease, or  proteinase K. Control binding represents binding of control-treated cells to the same tenascin substrate immediately before binding of  treated cells. Mean values for two experiments are shown with the range indicated by bars.
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Figure 1: Cell accumulation on plastic-adsorbed tenascin. (a) Ability of various cell types to accumulate on tenascin under flow. Cells were accumulated for 40 s each at 0.27 and 0.53 dynes/cm2. Cells were counted after the 0.53 dynes/cm2 accumulation step; all adherent cells were rolled. One of two representative experiments is shown with the range indicated by bars. (b) Effects of inhibitors and enzyme treatments on SKW3 cell accumulation on tenascin. SKW3 cells were treated for 10 min at 25°C with either 0.1% azide and 50 mM 2-deoxyglucose (Azide+DOG), inclusion of 5 mM EDTA or 10 mM EGTA and 1 mM Mg2+ in the assay, or treatment for 40 min at 37° with chondroitinase ABC, heparitinase, hyaluronidase, neuraminidase, neuraminidase and exo-β-galactosidase, O-glycoprotease, or proteinase K. Control binding represents binding of control-treated cells to the same tenascin substrate immediately before binding of treated cells. Mean values for two experiments are shown with the range indicated by bars.

Mentions: Freshly isolated peripheral blood lymphocytes and tonsillar lymphocytes introduced under flow adhered to plasticadsorbed tenascin at wall shear stresses of 0.27 and 0.53 dynes/cm2 (Fig. 1 a). In contrast, neutrophils from peripheral blood did not bind tenascin under flow. Adherent lymphocytes began rolling immediately after binding to the substrate and continued to roll as shear was increased. Binding to tenascin was threefold higher for tonsillar than for peripheral blood lymphocytes. Lymphocytes harvested from tonsil tissue contain activated cells that express the activation antigens CD26, CD30, CD39, CD69, CD70, and CD71 (8, 21, 58). Because cell lines are useful models for activated leukocytes, several cell lines were also evaluated for their ability to form rolling attachments to tenascin (Fig. 1 a). Most cell lines of T cell, B cell, and myeloid lineage bound tenascin, but the efficiency of binding varied widely. The SKW3 T lymphoblastoid cell line was chosen for subsequent studies because of its robust binding to tenascin under flow.


Tenascin supports lymphocyte rolling.

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

Cell accumulation on plastic-adsorbed tenascin. (a) Ability of various cell types to accumulate on tenascin under flow. Cells  were accumulated for 40 s each at 0.27 and 0.53 dynes/cm2. Cells were counted after the 0.53 dynes/cm2 accumulation step; all adherent  cells were rolled. One of two representative experiments is shown with the range indicated by bars. (b) Effects of inhibitors and enzyme  treatments on SKW3 cell accumulation on tenascin. SKW3 cells were treated for 10 min at 25°C with either 0.1% azide and 50 mM  2-deoxyglucose (Azide+DOG), inclusion of 5 mM EDTA or 10 mM EGTA and 1 mM Mg2+ in the assay, or treatment for 40 min at 37°  with chondroitinase ABC, heparitinase, hyaluronidase, neuraminidase, neuraminidase and exo-β-galactosidase, O-glycoprotease, or  proteinase K. Control binding represents binding of control-treated cells to the same tenascin substrate immediately before binding of  treated cells. Mean values for two experiments are shown with the range indicated by bars.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Cell accumulation on plastic-adsorbed tenascin. (a) Ability of various cell types to accumulate on tenascin under flow. Cells were accumulated for 40 s each at 0.27 and 0.53 dynes/cm2. Cells were counted after the 0.53 dynes/cm2 accumulation step; all adherent cells were rolled. One of two representative experiments is shown with the range indicated by bars. (b) Effects of inhibitors and enzyme treatments on SKW3 cell accumulation on tenascin. SKW3 cells were treated for 10 min at 25°C with either 0.1% azide and 50 mM 2-deoxyglucose (Azide+DOG), inclusion of 5 mM EDTA or 10 mM EGTA and 1 mM Mg2+ in the assay, or treatment for 40 min at 37° with chondroitinase ABC, heparitinase, hyaluronidase, neuraminidase, neuraminidase and exo-β-galactosidase, O-glycoprotease, or proteinase K. Control binding represents binding of control-treated cells to the same tenascin substrate immediately before binding of treated cells. Mean values for two experiments are shown with the range indicated by bars.
Mentions: Freshly isolated peripheral blood lymphocytes and tonsillar lymphocytes introduced under flow adhered to plasticadsorbed tenascin at wall shear stresses of 0.27 and 0.53 dynes/cm2 (Fig. 1 a). In contrast, neutrophils from peripheral blood did not bind tenascin under flow. Adherent lymphocytes began rolling immediately after binding to the substrate and continued to roll as shear was increased. Binding to tenascin was threefold higher for tonsillar than for peripheral blood lymphocytes. Lymphocytes harvested from tonsil tissue contain activated cells that express the activation antigens CD26, CD30, CD39, CD69, CD70, and CD71 (8, 21, 58). Because cell lines are useful models for activated leukocytes, several cell lines were also evaluated for their ability to form rolling attachments to tenascin (Fig. 1 a). Most cell lines of T cell, B cell, and myeloid lineage bound tenascin, but the efficiency of binding varied widely. The SKW3 T lymphoblastoid cell line was chosen for subsequent studies because of its robust binding to tenascin under flow.

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