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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appearance typical of LCs in naive skin, suggesting that the cells had detached from neighboring keratinocytes and withdrawn cellular processes in preparation for migration, but were unable to leave the epidermis.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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(A) Expression of α6 and β4 integrin subunits on LCs. Epidermal cell suspensions were prepared from the ears of naive mice by  treatment with 0.5% trypsin and LCs were identified by the expression of  MHC class II. The expression of integrin subunits was determined using  antibodies to α6 (GoH3 or EA-1) or β4 subunits in comparison with an  isotype-matched control antibody (IgG2a). Results show log fluorescence  (0–104 channels) for MHC class II (x axis) and integrin subunit (y axis).  The percentage of cells within each quadrant is given. (B) Expression of  α4, α6, and β4 integrin subunits on lymph node DCs. DCs were enriched  from the draining lymph nodes of oxazolone-treated mice by density gradient centrifugation on Metrizamide. DCs were distinguished from lymphocytes by forward scatter (FSC) versus side scatter (SSC) analysis and  identified by high expression of MHC class II and dual staining for  NLDC145 antigen. Histograms show the expression of α4, α6, and β4 integrin subunits on MHC class II positive DCs (solid lines) in comparison  with isotype-matched control antibodies (dashed lines). The profiles show  log fluorescence (0–104 channels) on the x axis and cell number (0–40) on  the y axis.
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Figure 1: (A) Expression of α6 and β4 integrin subunits on LCs. Epidermal cell suspensions were prepared from the ears of naive mice by treatment with 0.5% trypsin and LCs were identified by the expression of MHC class II. The expression of integrin subunits was determined using antibodies to α6 (GoH3 or EA-1) or β4 subunits in comparison with an isotype-matched control antibody (IgG2a). Results show log fluorescence (0–104 channels) for MHC class II (x axis) and integrin subunit (y axis). The percentage of cells within each quadrant is given. (B) Expression of α4, α6, and β4 integrin subunits on lymph node DCs. DCs were enriched from the draining lymph nodes of oxazolone-treated mice by density gradient centrifugation on Metrizamide. DCs were distinguished from lymphocytes by forward scatter (FSC) versus side scatter (SSC) analysis and identified by high expression of MHC class II and dual staining for NLDC145 antigen. Histograms show the expression of α4, α6, and β4 integrin subunits on MHC class II positive DCs (solid lines) in comparison with isotype-matched control antibodies (dashed lines). The profiles show log fluorescence (0–104 channels) on the x axis and cell number (0–40) on the y axis.

Mentions: We hypothesized that LCs may use distinct integrin adhesion receptors to interact with the underlying BM and other ECM proteins in skin and/or lymph nodes during migration from the skin to draining lymph nodes. We therefore examined epidermal LCs and lymph node DCs for the expression of integrins that mediate adhesion to two of the major components of BM and ECM, laminin and fibronectin (Fig. 1).


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

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

(A) Expression of α6 and β4 integrin subunits on LCs. Epidermal cell suspensions were prepared from the ears of naive mice by  treatment with 0.5% trypsin and LCs were identified by the expression of  MHC class II. The expression of integrin subunits was determined using  antibodies to α6 (GoH3 or EA-1) or β4 subunits in comparison with an  isotype-matched control antibody (IgG2a). Results show log fluorescence  (0–104 channels) for MHC class II (x axis) and integrin subunit (y axis).  The percentage of cells within each quadrant is given. (B) Expression of  α4, α6, and β4 integrin subunits on lymph node DCs. DCs were enriched  from the draining lymph nodes of oxazolone-treated mice by density gradient centrifugation on Metrizamide. DCs were distinguished from lymphocytes by forward scatter (FSC) versus side scatter (SSC) analysis and  identified by high expression of MHC class II and dual staining for  NLDC145 antigen. Histograms show the expression of α4, α6, and β4 integrin subunits on MHC class II positive DCs (solid lines) in comparison  with isotype-matched control antibodies (dashed lines). The profiles show  log fluorescence (0–104 channels) on the x axis and cell number (0–40) on  the y axis.
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Related In: Results  -  Collection

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Figure 1: (A) Expression of α6 and β4 integrin subunits on LCs. Epidermal cell suspensions were prepared from the ears of naive mice by treatment with 0.5% trypsin and LCs were identified by the expression of MHC class II. The expression of integrin subunits was determined using antibodies to α6 (GoH3 or EA-1) or β4 subunits in comparison with an isotype-matched control antibody (IgG2a). Results show log fluorescence (0–104 channels) for MHC class II (x axis) and integrin subunit (y axis). The percentage of cells within each quadrant is given. (B) Expression of α4, α6, and β4 integrin subunits on lymph node DCs. DCs were enriched from the draining lymph nodes of oxazolone-treated mice by density gradient centrifugation on Metrizamide. DCs were distinguished from lymphocytes by forward scatter (FSC) versus side scatter (SSC) analysis and identified by high expression of MHC class II and dual staining for NLDC145 antigen. Histograms show the expression of α4, α6, and β4 integrin subunits on MHC class II positive DCs (solid lines) in comparison with isotype-matched control antibodies (dashed lines). The profiles show log fluorescence (0–104 channels) on the x axis and cell number (0–40) on the y axis.
Mentions: We hypothesized that LCs may use distinct integrin adhesion receptors to interact with the underlying BM and other ECM proteins in skin and/or lymph nodes during migration from the skin to draining lymph nodes. We therefore examined epidermal LCs and lymph node DCs for the expression of integrins that mediate adhesion to two of the major components of BM and ECM, laminin and fibronectin (Fig. 1).

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