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P-selectin glycoprotein ligand-1 (PSGL-1) on T helper 1 but not on T helper 2 cells binds to P-selectin and supports migration into inflamed skin.

Borges E, Tietz W, Steegmaier M, Moll T, Hallmann R, Hamann A, Vestweber D - J. Exp. Med. (1997)

Bottom Line: PSGL-1 on Th2 cells, although expressed at similar levels as on Th1 cells, did not support binding to P-selectin.Thus, the P-selectin-binding form of PSGL-1 distinguishes Th1 cells from Th2 cells.Furthermore, PSGL-1 is relevant for the entry of Th1 cells into inflamed areas of the skin.

View Article: PubMed Central - PubMed

Affiliation: Institut für Zellbiologie, ZMBE, Universität Münster, Germany.

ABSTRACT
We have shown recently that mouse Th1 cells but not Th2 cells are selectively recruited into inflamed sites of a delayed-type hypersensitivity (DTH) reaction of the skin. This migration was blocked by monoclonal antibodies (mAb) against P- and E-selectin. Here we show that Th1 cells bind to P-selectin via the P-selectin glycoprotein ligand-1 (PSGL-1). This is the only glycoprotein ligand that was detectable by affinity isolation with a P-selectin-Ig fusion protein. Binding of Th1 cells to P-selectin, as analyzed by flow cytometry and in cell adhesion assays, was completely blocked by antibodies against PSGL-1. The same antibodies blocked partially the migration of Th1 cells into cutaneous DTH reactions. This blocking activity, in combination with that of a mAb against E-selectin, was additive. PSGL-1 on Th2 cells, although expressed at similar levels as on Th1 cells, did not support binding to P-selectin. Thus, the P-selectin-binding form of PSGL-1 distinguishes Th1 cells from Th2 cells. Furthermore, PSGL-1 is relevant for the entry of Th1 cells into inflamed areas of the skin. This is the first demonstration for the importance of PSGL-1 for mouse leukocyte recruitment in vivo.

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Adhesion of Th1 and Th2 cells to immobilized P-selectin–Ig.  Cell adhesion assays were performed with Th2 or Th1 cells (as indicated)  in 96-well microtiter plates coated with human IgG (hIgG) or P-selectin–  IgG (P-Sel). Before the assay, cells were incubated with HBSS(--), HBSS  with 50 μg/ml rabbit nonimmune antibodies (Co), or HBSS with 50 μg/ ml affinity-purified antibodies against PSGL-1 (α PSGL). Bound cells  were counted by computer-aided image analysis in three randomly chosen areas of defined size (per well) in three different wells for each determination. The depicted experiment represents one of three similar experiments.
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Figure 3: Adhesion of Th1 and Th2 cells to immobilized P-selectin–Ig. Cell adhesion assays were performed with Th2 or Th1 cells (as indicated) in 96-well microtiter plates coated with human IgG (hIgG) or P-selectin– IgG (P-Sel). Before the assay, cells were incubated with HBSS(--), HBSS with 50 μg/ml rabbit nonimmune antibodies (Co), or HBSS with 50 μg/ ml affinity-purified antibodies against PSGL-1 (α PSGL). Bound cells were counted by computer-aided image analysis in three randomly chosen areas of defined size (per well) in three different wells for each determination. The depicted experiment represents one of three similar experiments.

Mentions: Using nonstatic (rotation) adhesion assays, we analyzed the binding of Th1 and Th2 cells to P-selectin–Ig coated onto plastic. In agreement with the results obtained by FACS® analysis, Th1 cells bound efficiently to P-selectin– Ig and, this binding was blocked by 89% with affinity-purified antibodies against PSGL-1 (Fig. 3). Binding of Th2 cells to P-selectin was low and comparable to the residual binding of anti–PSGL-1–blocked Th1 cells. Thus, PSGL-1 almost exclusively mediates P-selectin binding to Th1 cells in FACS® analysis as well as in adhesion assays and is the only glycoprotein ligand that can be detected by affinity isolation with a P-selectin probe from these cells.


P-selectin glycoprotein ligand-1 (PSGL-1) on T helper 1 but not on T helper 2 cells binds to P-selectin and supports migration into inflamed skin.

Borges E, Tietz W, Steegmaier M, Moll T, Hallmann R, Hamann A, Vestweber D - J. Exp. Med. (1997)

Adhesion of Th1 and Th2 cells to immobilized P-selectin–Ig.  Cell adhesion assays were performed with Th2 or Th1 cells (as indicated)  in 96-well microtiter plates coated with human IgG (hIgG) or P-selectin–  IgG (P-Sel). Before the assay, cells were incubated with HBSS(--), HBSS  with 50 μg/ml rabbit nonimmune antibodies (Co), or HBSS with 50 μg/ ml affinity-purified antibodies against PSGL-1 (α PSGL). Bound cells  were counted by computer-aided image analysis in three randomly chosen areas of defined size (per well) in three different wells for each determination. The depicted experiment represents one of three similar experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196023&req=5

Figure 3: Adhesion of Th1 and Th2 cells to immobilized P-selectin–Ig. Cell adhesion assays were performed with Th2 or Th1 cells (as indicated) in 96-well microtiter plates coated with human IgG (hIgG) or P-selectin– IgG (P-Sel). Before the assay, cells were incubated with HBSS(--), HBSS with 50 μg/ml rabbit nonimmune antibodies (Co), or HBSS with 50 μg/ ml affinity-purified antibodies against PSGL-1 (α PSGL). Bound cells were counted by computer-aided image analysis in three randomly chosen areas of defined size (per well) in three different wells for each determination. The depicted experiment represents one of three similar experiments.
Mentions: Using nonstatic (rotation) adhesion assays, we analyzed the binding of Th1 and Th2 cells to P-selectin–Ig coated onto plastic. In agreement with the results obtained by FACS® analysis, Th1 cells bound efficiently to P-selectin– Ig and, this binding was blocked by 89% with affinity-purified antibodies against PSGL-1 (Fig. 3). Binding of Th2 cells to P-selectin was low and comparable to the residual binding of anti–PSGL-1–blocked Th1 cells. Thus, PSGL-1 almost exclusively mediates P-selectin binding to Th1 cells in FACS® analysis as well as in adhesion assays and is the only glycoprotein ligand that can be detected by affinity isolation with a P-selectin probe from these cells.

Bottom Line: PSGL-1 on Th2 cells, although expressed at similar levels as on Th1 cells, did not support binding to P-selectin.Thus, the P-selectin-binding form of PSGL-1 distinguishes Th1 cells from Th2 cells.Furthermore, PSGL-1 is relevant for the entry of Th1 cells into inflamed areas of the skin.

View Article: PubMed Central - PubMed

Affiliation: Institut für Zellbiologie, ZMBE, Universität Münster, Germany.

ABSTRACT
We have shown recently that mouse Th1 cells but not Th2 cells are selectively recruited into inflamed sites of a delayed-type hypersensitivity (DTH) reaction of the skin. This migration was blocked by monoclonal antibodies (mAb) against P- and E-selectin. Here we show that Th1 cells bind to P-selectin via the P-selectin glycoprotein ligand-1 (PSGL-1). This is the only glycoprotein ligand that was detectable by affinity isolation with a P-selectin-Ig fusion protein. Binding of Th1 cells to P-selectin, as analyzed by flow cytometry and in cell adhesion assays, was completely blocked by antibodies against PSGL-1. The same antibodies blocked partially the migration of Th1 cells into cutaneous DTH reactions. This blocking activity, in combination with that of a mAb against E-selectin, was additive. PSGL-1 on Th2 cells, although expressed at similar levels as on Th1 cells, did not support binding to P-selectin. Thus, the P-selectin-binding form of PSGL-1 distinguishes Th1 cells from Th2 cells. Furthermore, PSGL-1 is relevant for the entry of Th1 cells into inflamed areas of the skin. This is the first demonstration for the importance of PSGL-1 for mouse leukocyte recruitment in vivo.

Show MeSH
Related in: MedlinePlus