Limits...
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.

Show MeSH

Related in: MedlinePlus

FACS® analysis of  Th1 and Th2 cells with P-selectin–Ig and antibodies against  PSGL-1. Th1 and Th2 cells (as  indicated) were analyzed by flow  cytometry either with affinitypurified rabbit antibodies against  mouse PSGL-1  (A and B, solid  lines) or with P-selectin–Ig (C and  D, solid lines). Dotted lines show  negative control staining either  with nonimmune rabbit IgG (A  and B) or with human IgG (C  and D). E shows the staining of  Th1 cells with P-selectin–IgG  after preincubation of the cells  either with nonimmune rabbit  IgG (faint line) or with affinitypurified rabbit anti–PSGL-1 antibodies (bold line). P-selectin–Ig  was detected with PE-conjugated  F(ab′)2 donkey anti–human IgG,  and rabbit antibodies were detected with FITC-conjugated  goat anti–rabbit IgG.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196023&req=5

Figure 2: FACS® analysis of Th1 and Th2 cells with P-selectin–Ig and antibodies against PSGL-1. Th1 and Th2 cells (as indicated) were analyzed by flow cytometry either with affinitypurified rabbit antibodies against mouse PSGL-1 (A and B, solid lines) or with P-selectin–Ig (C and D, solid lines). Dotted lines show negative control staining either with nonimmune rabbit IgG (A and B) or with human IgG (C and D). E shows the staining of Th1 cells with P-selectin–IgG after preincubation of the cells either with nonimmune rabbit IgG (faint line) or with affinitypurified rabbit anti–PSGL-1 antibodies (bold line). P-selectin–Ig was detected with PE-conjugated F(ab′)2 donkey anti–human IgG, and rabbit antibodies were detected with FITC-conjugated goat anti–rabbit IgG.

Mentions: In line with the immunoprecipitation results, a comparison of Th1 and Th2 cells by FACS® analysis revealed that both cell types expressed similar levels of PSGL-1 (Fig. 2, A and B). However, 45% of the Th1 cells, but none of the Th2 cells, could be stained with P-selectin–Ig (Fig. 2, C and D). Staining with P-selectin–Ig was completely blocked by preincubating the cells with affinity-purified antibodies against PSGL-1, indicating that P-selectin binding to Th1 cells is mediated exclusively by PSGL-1 (Fig. 2 E).


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)

FACS® analysis of  Th1 and Th2 cells with P-selectin–Ig and antibodies against  PSGL-1. Th1 and Th2 cells (as  indicated) were analyzed by flow  cytometry either with affinitypurified rabbit antibodies against  mouse PSGL-1  (A and B, solid  lines) or with P-selectin–Ig (C and  D, solid lines). Dotted lines show  negative control staining either  with nonimmune rabbit IgG (A  and B) or with human IgG (C  and D). E shows the staining of  Th1 cells with P-selectin–IgG  after preincubation of the cells  either with nonimmune rabbit  IgG (faint line) or with affinitypurified rabbit anti–PSGL-1 antibodies (bold line). P-selectin–Ig  was detected with PE-conjugated  F(ab′)2 donkey anti–human IgG,  and rabbit antibodies were detected with FITC-conjugated  goat anti–rabbit IgG.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: FACS® analysis of Th1 and Th2 cells with P-selectin–Ig and antibodies against PSGL-1. Th1 and Th2 cells (as indicated) were analyzed by flow cytometry either with affinitypurified rabbit antibodies against mouse PSGL-1 (A and B, solid lines) or with P-selectin–Ig (C and D, solid lines). Dotted lines show negative control staining either with nonimmune rabbit IgG (A and B) or with human IgG (C and D). E shows the staining of Th1 cells with P-selectin–IgG after preincubation of the cells either with nonimmune rabbit IgG (faint line) or with affinitypurified rabbit anti–PSGL-1 antibodies (bold line). P-selectin–Ig was detected with PE-conjugated F(ab′)2 donkey anti–human IgG, and rabbit antibodies were detected with FITC-conjugated goat anti–rabbit IgG.
Mentions: In line with the immunoprecipitation results, a comparison of Th1 and Th2 cells by FACS® analysis revealed that both cell types expressed similar levels of PSGL-1 (Fig. 2, A and B). However, 45% of the Th1 cells, but none of the Th2 cells, could be stained with P-selectin–Ig (Fig. 2, C and D). Staining with P-selectin–Ig was completely blocked by preincubating the cells with affinity-purified antibodies against PSGL-1, indicating that P-selectin binding to Th1 cells is mediated exclusively by PSGL-1 (Fig. 2 E).

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