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Alpha 6 integrins are required for Langerhans cell migration from the epidermis.

Price AA, Cumberbatch M, Kimber I, Ager A - J. Exp. Med. (1997)

Bottom Line: RGD-containing peptides were also without effect on LC migration from skin explants.In contrast, alpha 4 integrins, or other integrin-dependent interactions with fibronectin that are mediated by the RGD recognition sequence, did not influence LC migration from the epidermis.In addition, alpha 4 integrins did not affect the accumulation of LCs as DCs in draining lymph nodes.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular Immunology, National Institute for Medical Research, London, United Kingdom.

ABSTRACT
Topical exposure of mice to chemical allergens results in the migration of epidermal Langerhans cells (LCs) from the skin and their accumulation as immunostimulatory dendritic cells (DCs) in draining lymph nodes. Epidermal cell-derived cytokines have been implicated in the maturation and migration of LCs, but the adhesion molecules that regulate LC migration have not been studied. We hypothesized that integrin-mediated interactions with extracellular matrix components of the skin and lymph node may regulate LC/DC migration. We found that alpha 6 integrins and alpha 4 integrins were differentially expressed by epidermal LCs and lymph node DCs. A majority of LCs (70%) expressed the alpha 6 integrin subunit, whereas DCs did not express alpha 6 integrins. In contrast, the alpha 4 integrin subunit was expressed at high levels on DCs but at much lower levels on LCs. The anti-alpha 6 integrin antibody, GoH3, which blocks binding to laminin, completely prevented the spontaneous migration of LCs from skin explants in vitro and the rapid migration of LCs from mouse ear skin induced after intradermal administration of TNF-alpha in vivo. GoH3 also reduced the accumulation of DCs in draining lymph nodes by a maximum of 70% after topical administration of the chemical allergen oxazolone. LCs remaining in the epidermis in the presence of GoH3 adopted a rounded morphology, rather than the interdigitating appearance typical of LCs in naive skin, suggesting that the cells had detached from neighboring keratinocytes and withdrawn cellular processes in preparation for migration, but were unable to leave the epidermis. The anti-alpha 4 integrin antibody PS/2, which blocks binding to fibronectin, had no effect on LC migration from the epidermis either in vitro or in vivo, or on the accumulation of DCs in draining lymph nodes after oxazolone application. RGD-containing peptides were also without effect on LC migration from skin explants. These results identify an important role for alpha 6 integrins in the migration of LC from the epidermis to the draining lymph node by regulating access across the epidermal basement membrane. In contrast, alpha 4 integrins, or other integrin-dependent interactions with fibronectin that are mediated by the RGD recognition sequence, did not influence LC migration from the epidermis. In addition, alpha 4 integrins did not affect the accumulation of LCs as DCs in draining lymph nodes.

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Immunofluorescence staining of epidermal LCs in situ. Groups of mice (n = 2) received a single 100 μl injection intraperitoneally of 40 μg  anti-α6 integrin antibody (GoH3) 2 h before intradermal injection into both ear pinnae of (A) 50 ng murine recombinant TNF-α or (B) 0.1% BSA alone.  Ears were removed 30 min later, epidermal sheets were prepared, and the morphology of LCs was assessed after indirect immunofluorescence staining for  MHC class II expression. Note the rounded morphology of LCs in TNF-α–treated ears in comparison with interdigitating morphology of LCs in controls. Magnification, ×800.
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Figure 6: Immunofluorescence staining of epidermal LCs in situ. Groups of mice (n = 2) received a single 100 μl injection intraperitoneally of 40 μg anti-α6 integrin antibody (GoH3) 2 h before intradermal injection into both ear pinnae of (A) 50 ng murine recombinant TNF-α or (B) 0.1% BSA alone. Ears were removed 30 min later, epidermal sheets were prepared, and the morphology of LCs was assessed after indirect immunofluorescence staining for MHC class II expression. Note the rounded morphology of LCs in TNF-α–treated ears in comparison with interdigitating morphology of LCs in controls. Magnification, ×800.

Mentions: To determine whether α6 integrins play a role in the migration of LCs from the epidermis in vivo, affinity-purified α6 integrin antibody was administered systemically and its effect on LC migration was determined after administration of mouse recombinant TNF-α (Fig. 5). In mice pretreated with 40 μg of isotype-matched control antibody (MAC 193) the number of LCs/mm2 was reduced from 826 ± 31 in naive animals to 671 ± 6 after administration of TNF-α, which is similar to previously reported results (8). However, mice pretreated with 40 μg anti-α6 integrin antibody GoH3 showed no reduction in the frequency of LCs; the number of LCs/mm2 at 833 ± 37 was similar to that in naive ear skin. The morphology of LCs remaining in the epidermis of α6 integrin antibody–treated mice was different to that of LCs in naive skin (Fig. 6). A proportion of LCs adopted a rounded morphology, similar to that of LCs in α6 integrin antibody–treated skin explants, as opposed to the interdigitating appearance typical of LCs. Systemic administration of GoH3 had no effect on the morphology of resident LCs in untreated skin.


Alpha 6 integrins are required for Langerhans cell migration from the epidermis.

Price AA, Cumberbatch M, Kimber I, Ager A - J. Exp. Med. (1997)

Immunofluorescence staining of epidermal LCs in situ. Groups of mice (n = 2) received a single 100 μl injection intraperitoneally of 40 μg  anti-α6 integrin antibody (GoH3) 2 h before intradermal injection into both ear pinnae of (A) 50 ng murine recombinant TNF-α or (B) 0.1% BSA alone.  Ears were removed 30 min later, epidermal sheets were prepared, and the morphology of LCs was assessed after indirect immunofluorescence staining for  MHC class II expression. Note the rounded morphology of LCs in TNF-α–treated ears in comparison with interdigitating morphology of LCs in controls. Magnification, ×800.
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Related In: Results  -  Collection

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Figure 6: Immunofluorescence staining of epidermal LCs in situ. Groups of mice (n = 2) received a single 100 μl injection intraperitoneally of 40 μg anti-α6 integrin antibody (GoH3) 2 h before intradermal injection into both ear pinnae of (A) 50 ng murine recombinant TNF-α or (B) 0.1% BSA alone. Ears were removed 30 min later, epidermal sheets were prepared, and the morphology of LCs was assessed after indirect immunofluorescence staining for MHC class II expression. Note the rounded morphology of LCs in TNF-α–treated ears in comparison with interdigitating morphology of LCs in controls. Magnification, ×800.
Mentions: To determine whether α6 integrins play a role in the migration of LCs from the epidermis in vivo, affinity-purified α6 integrin antibody was administered systemically and its effect on LC migration was determined after administration of mouse recombinant TNF-α (Fig. 5). In mice pretreated with 40 μg of isotype-matched control antibody (MAC 193) the number of LCs/mm2 was reduced from 826 ± 31 in naive animals to 671 ± 6 after administration of TNF-α, which is similar to previously reported results (8). However, mice pretreated with 40 μg anti-α6 integrin antibody GoH3 showed no reduction in the frequency of LCs; the number of LCs/mm2 at 833 ± 37 was similar to that in naive ear skin. The morphology of LCs remaining in the epidermis of α6 integrin antibody–treated mice was different to that of LCs in naive skin (Fig. 6). A proportion of LCs adopted a rounded morphology, similar to that of LCs in α6 integrin antibody–treated skin explants, as opposed to the interdigitating appearance typical of LCs. Systemic administration of GoH3 had no effect on the morphology of resident LCs in untreated skin.

Bottom Line: RGD-containing peptides were also without effect on LC migration from skin explants.In contrast, alpha 4 integrins, or other integrin-dependent interactions with fibronectin that are mediated by the RGD recognition sequence, did not influence LC migration from the epidermis.In addition, alpha 4 integrins did not affect the accumulation of LCs as DCs in draining lymph nodes.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular Immunology, National Institute for Medical Research, London, United Kingdom.

ABSTRACT
Topical exposure of mice to chemical allergens results in the migration of epidermal Langerhans cells (LCs) from the skin and their accumulation as immunostimulatory dendritic cells (DCs) in draining lymph nodes. Epidermal cell-derived cytokines have been implicated in the maturation and migration of LCs, but the adhesion molecules that regulate LC migration have not been studied. We hypothesized that integrin-mediated interactions with extracellular matrix components of the skin and lymph node may regulate LC/DC migration. We found that alpha 6 integrins and alpha 4 integrins were differentially expressed by epidermal LCs and lymph node DCs. A majority of LCs (70%) expressed the alpha 6 integrin subunit, whereas DCs did not express alpha 6 integrins. In contrast, the alpha 4 integrin subunit was expressed at high levels on DCs but at much lower levels on LCs. The anti-alpha 6 integrin antibody, GoH3, which blocks binding to laminin, completely prevented the spontaneous migration of LCs from skin explants in vitro and the rapid migration of LCs from mouse ear skin induced after intradermal administration of TNF-alpha in vivo. GoH3 also reduced the accumulation of DCs in draining lymph nodes by a maximum of 70% after topical administration of the chemical allergen oxazolone. LCs remaining in the epidermis in the presence of GoH3 adopted a rounded morphology, rather than the interdigitating appearance typical of LCs in naive skin, suggesting that the cells had detached from neighboring keratinocytes and withdrawn cellular processes in preparation for migration, but were unable to leave the epidermis. The anti-alpha 4 integrin antibody PS/2, which blocks binding to fibronectin, had no effect on LC migration from the epidermis either in vitro or in vivo, or on the accumulation of DCs in draining lymph nodes after oxazolone application. RGD-containing peptides were also without effect on LC migration from skin explants. These results identify an important role for alpha 6 integrins in the migration of LC from the epidermis to the draining lymph node by regulating access across the epidermal basement membrane. In contrast, alpha 4 integrins, or other integrin-dependent interactions with fibronectin that are mediated by the RGD recognition sequence, did not influence LC migration from the epidermis. In addition, alpha 4 integrins did not affect the accumulation of LCs as DCs in draining lymph nodes.

Show MeSH
Related in: MedlinePlus