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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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Comparison of KG1a cells rolling on tenascin and E-selectin. (a) Detachment profiles of KG1a cells on tenascin and two  plating dilutions of E-selectin. (b) Rolling velocities of KG1a  cells on tenascin and E-selectin. The mean values of two experiments are shown; ranges are indicated by bars.
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Figure 6: Comparison of KG1a cells rolling on tenascin and E-selectin. (a) Detachment profiles of KG1a cells on tenascin and two plating dilutions of E-selectin. (b) Rolling velocities of KG1a cells on tenascin and E-selectin. The mean values of two experiments are shown; ranges are indicated by bars.

Mentions: The characteristics of tenascin-mediated rolling were compared to rolling on E-selectin using KG1a cells, which bind to both tenascin and E-selectin (28). To directly compare the two adhesion systems, plating concentrations of E-selectin were varied until the detachment profile of KG1a cells on E-selectin was similar to that observed using a 50 μg/ml plating concentration of tenascin (Fig. 6 a). This required plating of purified E-selectin at concentrations 5- to 10-fold lower than the amount used routinely for experiments in our laboratory (42). At an E-selectin plating dilution of 1: 1,000, a level at which detachment from the two substrates was most similar, the E-selectin substrate was found to contain 34 sites/μm2, as measured by binding of radiolabeled mAb CL3. In contrast, the tenascin substrate contained 290 sites/μm2, as measured by binding of the function-blocking mAb M168. The tenascin substrate therefore had 8.5-fold more binding sites than the E-selectin substrate under conditions in which cells detached at roughly the same shear stresses.


Tenascin supports lymphocyte rolling.

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

Comparison of KG1a cells rolling on tenascin and E-selectin. (a) Detachment profiles of KG1a cells on tenascin and two  plating dilutions of E-selectin. (b) Rolling velocities of KG1a  cells on tenascin and E-selectin. The mean values of two experiments are shown; ranges are indicated by bars.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2139881&req=5

Figure 6: Comparison of KG1a cells rolling on tenascin and E-selectin. (a) Detachment profiles of KG1a cells on tenascin and two plating dilutions of E-selectin. (b) Rolling velocities of KG1a cells on tenascin and E-selectin. The mean values of two experiments are shown; ranges are indicated by bars.
Mentions: The characteristics of tenascin-mediated rolling were compared to rolling on E-selectin using KG1a cells, which bind to both tenascin and E-selectin (28). To directly compare the two adhesion systems, plating concentrations of E-selectin were varied until the detachment profile of KG1a cells on E-selectin was similar to that observed using a 50 μg/ml plating concentration of tenascin (Fig. 6 a). This required plating of purified E-selectin at concentrations 5- to 10-fold lower than the amount used routinely for experiments in our laboratory (42). At an E-selectin plating dilution of 1: 1,000, a level at which detachment from the two substrates was most similar, the E-selectin substrate was found to contain 34 sites/μm2, as measured by binding of radiolabeled mAb CL3. In contrast, the tenascin substrate contained 290 sites/μm2, as measured by binding of the function-blocking mAb M168. The tenascin substrate therefore had 8.5-fold more binding sites than the E-selectin substrate under conditions in which cells detached at roughly the same shear stresses.

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