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P-selectin and P-selectin glycoprotein ligand 1 are major determinants for Th1 cell recruitment to nonlymphoid effector sites in the intestinal lamina propria.

Haddad W, Cooper CJ, Zhang Z, Brown JB, Zhu Y, Issekutz A, Fuss I, Lee HO, Kansas GS, Barrett TA - J. Exp. Med. (2003)

Bottom Line: Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%.P-selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment.These data suggest that IL-12-induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medicine, Northwestern University Medical School, 745 N. Fairbanks, Searle 10-455, Chicago, IL 60611, USA.

ABSTRACT
The recruitment of activated T cell subsets to sites of effector immune responses is mediated by homing receptors induced upon activation in secondary lymphoid tissue. Using an adoptive transfer model, the intestinal recruitment of CD4+ T cells activated with intraperitoneal antigen in complete Freund's adjuvant was examined. The data demonstrate that activated CD4+ T cells recruited to intestinal Peyer's patches (PP) and lamina propria (LP) up-regulate functional P-selectin glycoprotein ligand 1 (PSGL-1). Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%. P-selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment. Studies of mice examined 3 d after adoptive transfer of differentiated T cell subsets revealed that Th1 but not Th2 cells were recruited to small intestine PP and LP. Mucosal addressin cell adhesion molecule blockade reduced Th1 recruitment to PP by 90% and to LP by >72%, whereas P-selectin blockade reduced Th1 recruitment to PP by 18% and Th1 recruitment to LP by 84%. These data suggest that IL-12-induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine.

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Related in: MedlinePlus

Anti–IL-12 inhibits P-selectin–Ig binding on activated T cells in MLN without effecting expression of homing receptors, α4β7, or L-selectin. (A) BALB/c mice pretreated with control mAb (C mAb) or anti–IL-12 were adoptively transferred with CD4+ KJ1–26.1+ T cells and challenged with i.p. OVA323–339/CFA. MLN cells were isolated from the mice 7d after Ag challenge and stained for P-selectin–Ig, α4β7, or L-selectin. Histograms of cells gated on the basis of CD4 and KJ1–26.1 staining are shown. Levels of control chimeric CD45-Ig staining are shown as the light line. Identical results were obtained with P-selectin–Ig stained in the presence of EDTA (not depicted). Numbers indicate the percentage of positively stained cells on the basis of control Ab staining. (B) Results of α4β7 and functional PSGL-1 expression on DO11.10 cells from pooled peripheral LN (PLN; brachial, axillary, and inguinal) taken from mice 3 d after i.p. OVA/CFA challenge, and stained with anti-α4β7 and P-selectin–Ig (Materials and Methods). Cells were gated on the basis of CD4 and KJ1–26.1 staining and numbers indicate the percentage of positively stained cells based on control Ab staining. (C) Results of α4β7 and functional PSGL-1 expression on MLN DO11.10 cells isolated from mice 2 d after i.p. OVA323–339 and LPS (OVA/LPS) challenge are shown. Cells were stained with anti-α4β7 and P-selectin–Ig (Materials and Methods) and gated on the basis of CD4 and KJ1–26.1 staining. Numbers indicate the percentage of positively stained cells based on control Ab staining. Data are representative of three independent experiments with similar results.
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fig2: Anti–IL-12 inhibits P-selectin–Ig binding on activated T cells in MLN without effecting expression of homing receptors, α4β7, or L-selectin. (A) BALB/c mice pretreated with control mAb (C mAb) or anti–IL-12 were adoptively transferred with CD4+ KJ1–26.1+ T cells and challenged with i.p. OVA323–339/CFA. MLN cells were isolated from the mice 7d after Ag challenge and stained for P-selectin–Ig, α4β7, or L-selectin. Histograms of cells gated on the basis of CD4 and KJ1–26.1 staining are shown. Levels of control chimeric CD45-Ig staining are shown as the light line. Identical results were obtained with P-selectin–Ig stained in the presence of EDTA (not depicted). Numbers indicate the percentage of positively stained cells on the basis of control Ab staining. (B) Results of α4β7 and functional PSGL-1 expression on DO11.10 cells from pooled peripheral LN (PLN; brachial, axillary, and inguinal) taken from mice 3 d after i.p. OVA/CFA challenge, and stained with anti-α4β7 and P-selectin–Ig (Materials and Methods). Cells were gated on the basis of CD4 and KJ1–26.1 staining and numbers indicate the percentage of positively stained cells based on control Ab staining. (C) Results of α4β7 and functional PSGL-1 expression on MLN DO11.10 cells isolated from mice 2 d after i.p. OVA323–339 and LPS (OVA/LPS) challenge are shown. Cells were stained with anti-α4β7 and P-selectin–Ig (Materials and Methods) and gated on the basis of CD4 and KJ1–26.1 staining. Numbers indicate the percentage of positively stained cells based on control Ab staining. Data are representative of three independent experiments with similar results.

Mentions: Anti–IL-12 mAb was purified from supernatant from clone 17.8 obtained from G. Trinchieri (Schering-Plough Research Institute, Dardilly, France; reference 18). Hermes-1 rat isotype control hybridoma was obtained at the Developmental Studies Hybridoma Bank. MECA-367 (anti-MAdCAM), 17.8, and Hermes-1 hybridomas were grown in a bioreactor (Heraeus) or as ascites and antibody purified with the Gammabind plus protein G column (Amersham Biosciences). The mouse E-selectin blocking mAb (9A9E3.F10, a rat IgG2b) and the mouse P-selectin blocking mAb (RMP-1, a mouse IgG2a) were gifts from A. Issekutz (Dalhousie University, Halifax, Nova Scotia, Canada; references 19, 20). Isotype control mouse IgG2a (clone G155–178) and rat IgG1 control (R3–34) were purchased from BD Biosciences. Purified antibodies for in vivo use were checked for endotoxin by the limulus assay (Associates of Cape Cod) and found to be <1 EU/ml. For mAb treatment, 2 mg anti–IL-12 or control mAb was given i.p. 2 h before Ag challenge. 0.2 mg anti-MAdCAM or control Ab was given daily in the indicated experiments. The anti–P-selectin, anti–E-selectin, or control mAbs was given 0.3 mg i.p. 2 h before Ag challenge, followed by 0.2 mg i.p on the third and fifth days after Ag challenge. In addition, a rat anti–mouse P-selectin mAb (RB40.34, rat IgG1) (a gift from K. Ley, University of Virginia, Charlottesville, VA) was used to confirm the result (see Fig. 2).


P-selectin and P-selectin glycoprotein ligand 1 are major determinants for Th1 cell recruitment to nonlymphoid effector sites in the intestinal lamina propria.

Haddad W, Cooper CJ, Zhang Z, Brown JB, Zhu Y, Issekutz A, Fuss I, Lee HO, Kansas GS, Barrett TA - J. Exp. Med. (2003)

Anti–IL-12 inhibits P-selectin–Ig binding on activated T cells in MLN without effecting expression of homing receptors, α4β7, or L-selectin. (A) BALB/c mice pretreated with control mAb (C mAb) or anti–IL-12 were adoptively transferred with CD4+ KJ1–26.1+ T cells and challenged with i.p. OVA323–339/CFA. MLN cells were isolated from the mice 7d after Ag challenge and stained for P-selectin–Ig, α4β7, or L-selectin. Histograms of cells gated on the basis of CD4 and KJ1–26.1 staining are shown. Levels of control chimeric CD45-Ig staining are shown as the light line. Identical results were obtained with P-selectin–Ig stained in the presence of EDTA (not depicted). Numbers indicate the percentage of positively stained cells on the basis of control Ab staining. (B) Results of α4β7 and functional PSGL-1 expression on DO11.10 cells from pooled peripheral LN (PLN; brachial, axillary, and inguinal) taken from mice 3 d after i.p. OVA/CFA challenge, and stained with anti-α4β7 and P-selectin–Ig (Materials and Methods). Cells were gated on the basis of CD4 and KJ1–26.1 staining and numbers indicate the percentage of positively stained cells based on control Ab staining. (C) Results of α4β7 and functional PSGL-1 expression on MLN DO11.10 cells isolated from mice 2 d after i.p. OVA323–339 and LPS (OVA/LPS) challenge are shown. Cells were stained with anti-α4β7 and P-selectin–Ig (Materials and Methods) and gated on the basis of CD4 and KJ1–26.1 staining. Numbers indicate the percentage of positively stained cells based on control Ab staining. Data are representative of three independent experiments with similar results.
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Related In: Results  -  Collection

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fig2: Anti–IL-12 inhibits P-selectin–Ig binding on activated T cells in MLN without effecting expression of homing receptors, α4β7, or L-selectin. (A) BALB/c mice pretreated with control mAb (C mAb) or anti–IL-12 were adoptively transferred with CD4+ KJ1–26.1+ T cells and challenged with i.p. OVA323–339/CFA. MLN cells were isolated from the mice 7d after Ag challenge and stained for P-selectin–Ig, α4β7, or L-selectin. Histograms of cells gated on the basis of CD4 and KJ1–26.1 staining are shown. Levels of control chimeric CD45-Ig staining are shown as the light line. Identical results were obtained with P-selectin–Ig stained in the presence of EDTA (not depicted). Numbers indicate the percentage of positively stained cells on the basis of control Ab staining. (B) Results of α4β7 and functional PSGL-1 expression on DO11.10 cells from pooled peripheral LN (PLN; brachial, axillary, and inguinal) taken from mice 3 d after i.p. OVA/CFA challenge, and stained with anti-α4β7 and P-selectin–Ig (Materials and Methods). Cells were gated on the basis of CD4 and KJ1–26.1 staining and numbers indicate the percentage of positively stained cells based on control Ab staining. (C) Results of α4β7 and functional PSGL-1 expression on MLN DO11.10 cells isolated from mice 2 d after i.p. OVA323–339 and LPS (OVA/LPS) challenge are shown. Cells were stained with anti-α4β7 and P-selectin–Ig (Materials and Methods) and gated on the basis of CD4 and KJ1–26.1 staining. Numbers indicate the percentage of positively stained cells based on control Ab staining. Data are representative of three independent experiments with similar results.
Mentions: Anti–IL-12 mAb was purified from supernatant from clone 17.8 obtained from G. Trinchieri (Schering-Plough Research Institute, Dardilly, France; reference 18). Hermes-1 rat isotype control hybridoma was obtained at the Developmental Studies Hybridoma Bank. MECA-367 (anti-MAdCAM), 17.8, and Hermes-1 hybridomas were grown in a bioreactor (Heraeus) or as ascites and antibody purified with the Gammabind plus protein G column (Amersham Biosciences). The mouse E-selectin blocking mAb (9A9E3.F10, a rat IgG2b) and the mouse P-selectin blocking mAb (RMP-1, a mouse IgG2a) were gifts from A. Issekutz (Dalhousie University, Halifax, Nova Scotia, Canada; references 19, 20). Isotype control mouse IgG2a (clone G155–178) and rat IgG1 control (R3–34) were purchased from BD Biosciences. Purified antibodies for in vivo use were checked for endotoxin by the limulus assay (Associates of Cape Cod) and found to be <1 EU/ml. For mAb treatment, 2 mg anti–IL-12 or control mAb was given i.p. 2 h before Ag challenge. 0.2 mg anti-MAdCAM or control Ab was given daily in the indicated experiments. The anti–P-selectin, anti–E-selectin, or control mAbs was given 0.3 mg i.p. 2 h before Ag challenge, followed by 0.2 mg i.p on the third and fifth days after Ag challenge. In addition, a rat anti–mouse P-selectin mAb (RB40.34, rat IgG1) (a gift from K. Ley, University of Virginia, Charlottesville, VA) was used to confirm the result (see Fig. 2).

Bottom Line: Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%.P-selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment.These data suggest that IL-12-induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Medicine, Northwestern University Medical School, 745 N. Fairbanks, Searle 10-455, Chicago, IL 60611, USA.

ABSTRACT
The recruitment of activated T cell subsets to sites of effector immune responses is mediated by homing receptors induced upon activation in secondary lymphoid tissue. Using an adoptive transfer model, the intestinal recruitment of CD4+ T cells activated with intraperitoneal antigen in complete Freund's adjuvant was examined. The data demonstrate that activated CD4+ T cells recruited to intestinal Peyer's patches (PP) and lamina propria (LP) up-regulate functional P-selectin glycoprotein ligand 1 (PSGL-1). Blockade of IL-12 inhibited functional PSGL-1 expression and reduced PP and LP CD4+ T cell recruitment by >40%. P-selectin blockade reduced LP recruitment of activated cells by 56% without affecting PP recruitment. Studies of mice examined 3 d after adoptive transfer of differentiated T cell subsets revealed that Th1 but not Th2 cells were recruited to small intestine PP and LP. Mucosal addressin cell adhesion molecule blockade reduced Th1 recruitment to PP by 90% and to LP by >72%, whereas P-selectin blockade reduced Th1 recruitment to PP by 18% and Th1 recruitment to LP by 84%. These data suggest that IL-12-induced functional PSGL-1 expression is a major determinant for the recruitment of Th1 effector cells to noninflamed as well as inflamed intestine.

Show MeSH
Related in: MedlinePlus