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Chemokine receptor CCR9 contributes to the localization of plasma cells to the small intestine.

Pabst O, Ohl L, Wendland M, Wurbel MA, Kremmer E, Malissen B, Förster R - J. Exp. Med. (2004)

Bottom Line: Humoral immunity in the gut-associated lymphoid tissue is characterized by the production of immunoglobulin A (IgA) by antibody-secreting plasma cells (PCs) in the lamina propria.In CCR9-deficient mice, IgA+ PCs are substantially reduced in number in the lamina propria of the small intestine.These findings provide profound in vivo evidence that CCL25/CCR9 guides PCs into the small intestine.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.

ABSTRACT
Humoral immunity in the gut-associated lymphoid tissue is characterized by the production of immunoglobulin A (IgA) by antibody-secreting plasma cells (PCs) in the lamina propria. The chemokine CCL25 is expressed by intestinal epithelial cells and is capable of inducing chemotaxis of IgA+ PCs in vitro. Using a newly generated monoclonal antibody against murine CCR9, we show that IgA+ PCs express high levels of CCR9 in the mesenteric lymph node (MLN) and Peyer's patches (PPs), but down-regulate CCR9 once they are located in the small intestine. In CCR9-deficient mice, IgA+ PCs are substantially reduced in number in the lamina propria of the small intestine. In adoptive transfer experiments, CCR9-deficient IgA+ PCs show reduced migration into the small intestine compared with wild-type controls. Furthermore, CCR9 mutants fail to mount a regular IgA response to an orally administered antigen, although the architecture and cell type composition of PPs and MLN are unaffected and are functional for the generation of IgA PCs. These findings provide profound in vivo evidence that CCL25/CCR9 guides PCs into the small intestine.

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Regular generation of IgA+ PCs in CCR9-deficient animals. Wild-type (A and C) and CCR9-deficient mice (B and D) were gavaged with fluorescent latex beads and treated after 24 h with PBS alone (A and B) or CT in PBS (C and D). After 24 h, PP were removed, sectioned, and stained for CD11c (red), CD3 (green), and nuclei (blue). Wild-type and CCR9-deficient mice showed no differences in architecture, nor mobilization of CD11c+ cells or mobilization of bead-loaded cells. (A, inset) Arrows point to bead-loaded CD11c+ cells. Sections of MLN of wild type (E) and CCR9 mutants (F) that had been immunized with OVA and CT five times at 10-d intervals were stained with anti-IgA (green), anti-IgM (red), and anti-CD3 (blue). Similar numbers of IgA+ PCs are present in wild type and CCR9 MLN, demonstrating that the generation of IgA PCs is not grossly altered in CCR9-deficient mice.
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fig4: Regular generation of IgA+ PCs in CCR9-deficient animals. Wild-type (A and C) and CCR9-deficient mice (B and D) were gavaged with fluorescent latex beads and treated after 24 h with PBS alone (A and B) or CT in PBS (C and D). After 24 h, PP were removed, sectioned, and stained for CD11c (red), CD3 (green), and nuclei (blue). Wild-type and CCR9-deficient mice showed no differences in architecture, nor mobilization of CD11c+ cells or mobilization of bead-loaded cells. (A, inset) Arrows point to bead-loaded CD11c+ cells. Sections of MLN of wild type (E) and CCR9 mutants (F) that had been immunized with OVA and CT five times at 10-d intervals were stained with anti-IgA (green), anti-IgM (red), and anti-CD3 (blue). Similar numbers of IgA+ PCs are present in wild type and CCR9 MLN, demonstrating that the generation of IgA PCs is not grossly altered in CCR9-deficient mice.

Mentions: Because it is assumed that the induction of an IgA-specific antibody response after oral application of antigen plus CT requires antigen presentation within morphologically intact PPs and MLN, we further analyzed both organs. We used immunohistology and flow cytometry to identify possible alterations in cellular composition or architecture of both organs in CCR9 mutants that contained normal numbers of B and T cells, and both cell types were located in their appropriate microenvironments. In addition, PPs contained normal numbers of CD11c+, CD11b−, and CD11c+CD11b+ DCs (unpublished data). Recently, it has been described that DCs of the SED can be labeled and their path subsequently followed using fluorescent latex beads (4). Because we found CCR9-expressing cells, including DC within the SED (unpublished data), we tested whether the mobilization of DCs of the SED is affected in CCR9 mutants. Wild-type and CCR9-deficient mice were deprived of water and food for 4 h and subsequently gavaged with 1012 fluorescent latex beads (200 nm diameter; Polysciences) per animal. After 24 h, the mice were gavaged with 50 μg of CT. After another 24 h, PPs and MLN were sectioned and stained for CD11c and CD3 (Fig. 4, A–D, PPs only). In wild type and CCR9 mutants, application of CT lead to reduced numbers of CD11c+ cells (red) present in the SED (Fig. 4, A–D). To quantify this effect, the absolute number of fluorescent beads in the SED was counted in 20 PPs derived from four wild type and four CCR9 mutants. In both wild type and CCR9 mutants, CT administration triggers an identical fivefold decrease of bead-labeled DCs localized to the SED, suggesting that no differences in DC mobilization exist and that the activation of naive B cells should proceed normally in these CCR9−/− organs (unpublished data). Indeed, analysis of cryosections from MLN of wild-type and mutant mice that were immunized five times with OVA and CT revealed comparable numbers of IgA+ PCs in both strains (Fig. 4, E and F). Furthermore, in wild type and mutants likewise, the total numbers of IgA+ PCs strongly increased after immunization in comparison to nonimmunized mice (unpublished data) strongly supporting the idea that the generation of IgA PCs is not affected by CCR9 deficiency.


Chemokine receptor CCR9 contributes to the localization of plasma cells to the small intestine.

Pabst O, Ohl L, Wendland M, Wurbel MA, Kremmer E, Malissen B, Förster R - J. Exp. Med. (2004)

Regular generation of IgA+ PCs in CCR9-deficient animals. Wild-type (A and C) and CCR9-deficient mice (B and D) were gavaged with fluorescent latex beads and treated after 24 h with PBS alone (A and B) or CT in PBS (C and D). After 24 h, PP were removed, sectioned, and stained for CD11c (red), CD3 (green), and nuclei (blue). Wild-type and CCR9-deficient mice showed no differences in architecture, nor mobilization of CD11c+ cells or mobilization of bead-loaded cells. (A, inset) Arrows point to bead-loaded CD11c+ cells. Sections of MLN of wild type (E) and CCR9 mutants (F) that had been immunized with OVA and CT five times at 10-d intervals were stained with anti-IgA (green), anti-IgM (red), and anti-CD3 (blue). Similar numbers of IgA+ PCs are present in wild type and CCR9 MLN, demonstrating that the generation of IgA PCs is not grossly altered in CCR9-deficient mice.
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Related In: Results  -  Collection

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fig4: Regular generation of IgA+ PCs in CCR9-deficient animals. Wild-type (A and C) and CCR9-deficient mice (B and D) were gavaged with fluorescent latex beads and treated after 24 h with PBS alone (A and B) or CT in PBS (C and D). After 24 h, PP were removed, sectioned, and stained for CD11c (red), CD3 (green), and nuclei (blue). Wild-type and CCR9-deficient mice showed no differences in architecture, nor mobilization of CD11c+ cells or mobilization of bead-loaded cells. (A, inset) Arrows point to bead-loaded CD11c+ cells. Sections of MLN of wild type (E) and CCR9 mutants (F) that had been immunized with OVA and CT five times at 10-d intervals were stained with anti-IgA (green), anti-IgM (red), and anti-CD3 (blue). Similar numbers of IgA+ PCs are present in wild type and CCR9 MLN, demonstrating that the generation of IgA PCs is not grossly altered in CCR9-deficient mice.
Mentions: Because it is assumed that the induction of an IgA-specific antibody response after oral application of antigen plus CT requires antigen presentation within morphologically intact PPs and MLN, we further analyzed both organs. We used immunohistology and flow cytometry to identify possible alterations in cellular composition or architecture of both organs in CCR9 mutants that contained normal numbers of B and T cells, and both cell types were located in their appropriate microenvironments. In addition, PPs contained normal numbers of CD11c+, CD11b−, and CD11c+CD11b+ DCs (unpublished data). Recently, it has been described that DCs of the SED can be labeled and their path subsequently followed using fluorescent latex beads (4). Because we found CCR9-expressing cells, including DC within the SED (unpublished data), we tested whether the mobilization of DCs of the SED is affected in CCR9 mutants. Wild-type and CCR9-deficient mice were deprived of water and food for 4 h and subsequently gavaged with 1012 fluorescent latex beads (200 nm diameter; Polysciences) per animal. After 24 h, the mice were gavaged with 50 μg of CT. After another 24 h, PPs and MLN were sectioned and stained for CD11c and CD3 (Fig. 4, A–D, PPs only). In wild type and CCR9 mutants, application of CT lead to reduced numbers of CD11c+ cells (red) present in the SED (Fig. 4, A–D). To quantify this effect, the absolute number of fluorescent beads in the SED was counted in 20 PPs derived from four wild type and four CCR9 mutants. In both wild type and CCR9 mutants, CT administration triggers an identical fivefold decrease of bead-labeled DCs localized to the SED, suggesting that no differences in DC mobilization exist and that the activation of naive B cells should proceed normally in these CCR9−/− organs (unpublished data). Indeed, analysis of cryosections from MLN of wild-type and mutant mice that were immunized five times with OVA and CT revealed comparable numbers of IgA+ PCs in both strains (Fig. 4, E and F). Furthermore, in wild type and mutants likewise, the total numbers of IgA+ PCs strongly increased after immunization in comparison to nonimmunized mice (unpublished data) strongly supporting the idea that the generation of IgA PCs is not affected by CCR9 deficiency.

Bottom Line: Humoral immunity in the gut-associated lymphoid tissue is characterized by the production of immunoglobulin A (IgA) by antibody-secreting plasma cells (PCs) in the lamina propria.In CCR9-deficient mice, IgA+ PCs are substantially reduced in number in the lamina propria of the small intestine.These findings provide profound in vivo evidence that CCL25/CCR9 guides PCs into the small intestine.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.

ABSTRACT
Humoral immunity in the gut-associated lymphoid tissue is characterized by the production of immunoglobulin A (IgA) by antibody-secreting plasma cells (PCs) in the lamina propria. The chemokine CCL25 is expressed by intestinal epithelial cells and is capable of inducing chemotaxis of IgA+ PCs in vitro. Using a newly generated monoclonal antibody against murine CCR9, we show that IgA+ PCs express high levels of CCR9 in the mesenteric lymph node (MLN) and Peyer's patches (PPs), but down-regulate CCR9 once they are located in the small intestine. In CCR9-deficient mice, IgA+ PCs are substantially reduced in number in the lamina propria of the small intestine. In adoptive transfer experiments, CCR9-deficient IgA+ PCs show reduced migration into the small intestine compared with wild-type controls. Furthermore, CCR9 mutants fail to mount a regular IgA response to an orally administered antigen, although the architecture and cell type composition of PPs and MLN are unaffected and are functional for the generation of IgA PCs. These findings provide profound in vivo evidence that CCL25/CCR9 guides PCs into the small intestine.

Show MeSH