Limits...
Accumulation of immature Langerhans cells in human lymph nodes draining chronically inflamed skin.

Geissmann F, Dieu-Nosjean MC, Dezutter C, Valladeau J, Kayal S, Leborgne M, Brousse N, Saeland S, Davoust J - J. Exp. Med. (2002)

Bottom Line: This indicates that LC migration and maturation can be independently regulated events.We suggest that during DL, inflammatory stimuli in the skin increase the migration of LCs to the lymph node but without associated maturation.Immature LCs might regulate immune responses during chronic inflammation.

View Article: PubMed Central - PubMed

Affiliation: Service d'Anatomie Pathologique, IFR Necker-Enfants Malades, 75015 Paris, France. geissman@necker.fr

ABSTRACT
The coordinated migration and maturation of dendritic cells (DCs) such as intraepithelial Langerhans cells (LCs) is considered critical for T cell priming in response to inflammation in the periphery. However, little is known about the role of inflammatory mediators for LC maturation and recruitment to lymph nodes in vivo. Here we show in human dermatopathic lymphadenitis (DL), which features an expanded population of LCs in one draining lymph node associated with inflammatory lesions in its tributary skin area, that the Langerin/CD207(+) LCs constitute a predominant population of immature DCs, which express CD1a, and CD68, but not CD83, CD86, and DC-lysosomal-associated membrane protein (LAMP)/CD208. Using LC-type cells generated in vitro in the presence of transforming growth factor (TGF)-beta1, we further found that tumor necrosis factor (TNF)-alpha, as a prototype proinflammatory factor, and a variety of inflammatory stimuli and bacterial products, increase Langerin expression and Langerin dependent Birbeck granules formation in cell which nevertheless lack costimulatory molecules, DC-LAMP/CD208 and potent T cell stimulatory activity but express CCR7 and respond to the lymph node homing chemokines CCL19 and CCL21. This indicates that LC migration and maturation can be independently regulated events. We suggest that during DL, inflammatory stimuli in the skin increase the migration of LCs to the lymph node but without associated maturation. Immature LCs might regulate immune responses during chronic inflammation.

Show MeSH

Related in: MedlinePlus

LC-type DCs cultured with TNF-α express CCR7 and migrate toward CCR7 ligands but are not functionally mature. (A) Autologous antigen presentation. LC-type cells (left) and DCi (right) unstimulated (open circles), or treated with TNF-α (closed circles) or CD40-activated LCs (closed triangles) were pulsed with TT. T cell proliferation was measured as indicated in Materials and Methods. Results are expressed as mean of triplicates in a representative experiment. SD were <20%. (B) Both SLC and ELC trigger migration of TNF-α and E. coli treated, but not control unstimulated LC-type cells. Day 6, LCs were treated for 48 h with TNF-α (10 ng/ml) or with heat-inactivated E. coli (50:1 ratio, right). Then, samples were recovered and cells were tested for their capacity to migrate in response to ELC (100 ng/ml) and to SLC (100 ng/ml). Migration assays were performed in Boyden microchambers. (Left) Results are expressed as number of migrating LCs per two low power fields (original magnification: ×20). (Right) Results are expressed as migration index compared with medium alone (without chemo-kine). Results are representative of more than three independent experiments. (C) CCR7 expression. Day 6, LC-type and DC-type cells were treated for 48 h with 10 ng/ml TNF-α. Cells were washed and stained with PE-conjugated anti-Langerin, FITC-anti-DR and anti-CCR7 or with isotype-matched controls antibodies and analyzed by flow cytometry. In the left panel, the dot-plot represent langerin and HLA-DR expression of LC-type cells treated with TNF-α. On the center and right panels histograms represent CCR7 expression on gated-cell populations (R1:Langerin1 DRlo; R2: Langerin-, DRhi). Thick lines represent the labeling obtained with CCR7 antibody and thin lines represent labeling with the isotype-matched control. Results are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196053&req=5

fig5: LC-type DCs cultured with TNF-α express CCR7 and migrate toward CCR7 ligands but are not functionally mature. (A) Autologous antigen presentation. LC-type cells (left) and DCi (right) unstimulated (open circles), or treated with TNF-α (closed circles) or CD40-activated LCs (closed triangles) were pulsed with TT. T cell proliferation was measured as indicated in Materials and Methods. Results are expressed as mean of triplicates in a representative experiment. SD were <20%. (B) Both SLC and ELC trigger migration of TNF-α and E. coli treated, but not control unstimulated LC-type cells. Day 6, LCs were treated for 48 h with TNF-α (10 ng/ml) or with heat-inactivated E. coli (50:1 ratio, right). Then, samples were recovered and cells were tested for their capacity to migrate in response to ELC (100 ng/ml) and to SLC (100 ng/ml). Migration assays were performed in Boyden microchambers. (Left) Results are expressed as number of migrating LCs per two low power fields (original magnification: ×20). (Right) Results are expressed as migration index compared with medium alone (without chemo-kine). Results are representative of more than three independent experiments. (C) CCR7 expression. Day 6, LC-type and DC-type cells were treated for 48 h with 10 ng/ml TNF-α. Cells were washed and stained with PE-conjugated anti-Langerin, FITC-anti-DR and anti-CCR7 or with isotype-matched controls antibodies and analyzed by flow cytometry. In the left panel, the dot-plot represent langerin and HLA-DR expression of LC-type cells treated with TNF-α. On the center and right panels histograms represent CCR7 expression on gated-cell populations (R1:Langerin1 DRlo; R2: Langerin-, DRhi). Thick lines represent the labeling obtained with CCR7 antibody and thin lines represent labeling with the isotype-matched control. Results are representative of two independent experiments.

Mentions: Next, we investigated other functional properties of LC-type cells activated by TNF-α such as their Ag presentation capacity and their ability to migrate toward the LN environment. First, in accordance with our previous results (21) we found that treatment with TNF-α did not enable LC-type cells to stimulate the proliferation of autologous antigen specific memory T cells in vitro (Fig. 5 A), confirming the immature state of the TNF-α–treated LC-type cells. In contrast, monocyte-derived DCi generated in the absence of TGF-β1 became mature after exposure to TNF-α and thereafter, efficiently stimulated autologous antigen specific T cells (Fig. 5 A).


Accumulation of immature Langerhans cells in human lymph nodes draining chronically inflamed skin.

Geissmann F, Dieu-Nosjean MC, Dezutter C, Valladeau J, Kayal S, Leborgne M, Brousse N, Saeland S, Davoust J - J. Exp. Med. (2002)

LC-type DCs cultured with TNF-α express CCR7 and migrate toward CCR7 ligands but are not functionally mature. (A) Autologous antigen presentation. LC-type cells (left) and DCi (right) unstimulated (open circles), or treated with TNF-α (closed circles) or CD40-activated LCs (closed triangles) were pulsed with TT. T cell proliferation was measured as indicated in Materials and Methods. Results are expressed as mean of triplicates in a representative experiment. SD were <20%. (B) Both SLC and ELC trigger migration of TNF-α and E. coli treated, but not control unstimulated LC-type cells. Day 6, LCs were treated for 48 h with TNF-α (10 ng/ml) or with heat-inactivated E. coli (50:1 ratio, right). Then, samples were recovered and cells were tested for their capacity to migrate in response to ELC (100 ng/ml) and to SLC (100 ng/ml). Migration assays were performed in Boyden microchambers. (Left) Results are expressed as number of migrating LCs per two low power fields (original magnification: ×20). (Right) Results are expressed as migration index compared with medium alone (without chemo-kine). Results are representative of more than three independent experiments. (C) CCR7 expression. Day 6, LC-type and DC-type cells were treated for 48 h with 10 ng/ml TNF-α. Cells were washed and stained with PE-conjugated anti-Langerin, FITC-anti-DR and anti-CCR7 or with isotype-matched controls antibodies and analyzed by flow cytometry. In the left panel, the dot-plot represent langerin and HLA-DR expression of LC-type cells treated with TNF-α. On the center and right panels histograms represent CCR7 expression on gated-cell populations (R1:Langerin1 DRlo; R2: Langerin-, DRhi). Thick lines represent the labeling obtained with CCR7 antibody and thin lines represent labeling with the isotype-matched control. Results are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196053&req=5

fig5: LC-type DCs cultured with TNF-α express CCR7 and migrate toward CCR7 ligands but are not functionally mature. (A) Autologous antigen presentation. LC-type cells (left) and DCi (right) unstimulated (open circles), or treated with TNF-α (closed circles) or CD40-activated LCs (closed triangles) were pulsed with TT. T cell proliferation was measured as indicated in Materials and Methods. Results are expressed as mean of triplicates in a representative experiment. SD were <20%. (B) Both SLC and ELC trigger migration of TNF-α and E. coli treated, but not control unstimulated LC-type cells. Day 6, LCs were treated for 48 h with TNF-α (10 ng/ml) or with heat-inactivated E. coli (50:1 ratio, right). Then, samples were recovered and cells were tested for their capacity to migrate in response to ELC (100 ng/ml) and to SLC (100 ng/ml). Migration assays were performed in Boyden microchambers. (Left) Results are expressed as number of migrating LCs per two low power fields (original magnification: ×20). (Right) Results are expressed as migration index compared with medium alone (without chemo-kine). Results are representative of more than three independent experiments. (C) CCR7 expression. Day 6, LC-type and DC-type cells were treated for 48 h with 10 ng/ml TNF-α. Cells were washed and stained with PE-conjugated anti-Langerin, FITC-anti-DR and anti-CCR7 or with isotype-matched controls antibodies and analyzed by flow cytometry. In the left panel, the dot-plot represent langerin and HLA-DR expression of LC-type cells treated with TNF-α. On the center and right panels histograms represent CCR7 expression on gated-cell populations (R1:Langerin1 DRlo; R2: Langerin-, DRhi). Thick lines represent the labeling obtained with CCR7 antibody and thin lines represent labeling with the isotype-matched control. Results are representative of two independent experiments.
Mentions: Next, we investigated other functional properties of LC-type cells activated by TNF-α such as their Ag presentation capacity and their ability to migrate toward the LN environment. First, in accordance with our previous results (21) we found that treatment with TNF-α did not enable LC-type cells to stimulate the proliferation of autologous antigen specific memory T cells in vitro (Fig. 5 A), confirming the immature state of the TNF-α–treated LC-type cells. In contrast, monocyte-derived DCi generated in the absence of TGF-β1 became mature after exposure to TNF-α and thereafter, efficiently stimulated autologous antigen specific T cells (Fig. 5 A).

Bottom Line: This indicates that LC migration and maturation can be independently regulated events.We suggest that during DL, inflammatory stimuli in the skin increase the migration of LCs to the lymph node but without associated maturation.Immature LCs might regulate immune responses during chronic inflammation.

View Article: PubMed Central - PubMed

Affiliation: Service d'Anatomie Pathologique, IFR Necker-Enfants Malades, 75015 Paris, France. geissman@necker.fr

ABSTRACT
The coordinated migration and maturation of dendritic cells (DCs) such as intraepithelial Langerhans cells (LCs) is considered critical for T cell priming in response to inflammation in the periphery. However, little is known about the role of inflammatory mediators for LC maturation and recruitment to lymph nodes in vivo. Here we show in human dermatopathic lymphadenitis (DL), which features an expanded population of LCs in one draining lymph node associated with inflammatory lesions in its tributary skin area, that the Langerin/CD207(+) LCs constitute a predominant population of immature DCs, which express CD1a, and CD68, but not CD83, CD86, and DC-lysosomal-associated membrane protein (LAMP)/CD208. Using LC-type cells generated in vitro in the presence of transforming growth factor (TGF)-beta1, we further found that tumor necrosis factor (TNF)-alpha, as a prototype proinflammatory factor, and a variety of inflammatory stimuli and bacterial products, increase Langerin expression and Langerin dependent Birbeck granules formation in cell which nevertheless lack costimulatory molecules, DC-LAMP/CD208 and potent T cell stimulatory activity but express CCR7 and respond to the lymph node homing chemokines CCL19 and CCL21. This indicates that LC migration and maturation can be independently regulated events. We suggest that during DL, inflammatory stimuli in the skin increase the migration of LCs to the lymph node but without associated maturation. Immature LCs might regulate immune responses during chronic inflammation.

Show MeSH
Related in: MedlinePlus