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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appearance typical of LCs in naive skin, suggesting that the cells had detached from neighboring keratinocytes and withdrawn cellular processes in preparation for migration, but were unable to leave the epidermis.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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The effect of (A) anti-α6  and (B) anti-α4 integrin antibodies on  oxazolone-induced DC accumulation in lymph nodes in vivo. Antibody was administered to groups of  10 mice in single 100 μl injections  intraperitoneally for the 10, 20, and  200 μg doses or in single 30 μl injections intradermally into the dorsum  of both ears for 12 μg dose. The 200  μg dose of anti-α4 integrin antibody  was given in 2 × 100 μg doses 2 h  before and 8 h after oxazolone treatment. Each graph represents a separate experiment and the amount of antibody received per mouse, and the method of administration is indicated below  each graph. 2 h after antibody administration, mice received 25 μl of 0.5% oxazolone on the dorsum of both ears. Draining auricular lymph nodes were  excised 18 h later and the number of DCs per lymph node was determined. Results for isotype-matched control antibody (hatched bars) and integrin antibody (solid bars) are compared with untreated, naive mice (open bars).
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Figure 7: The effect of (A) anti-α6 and (B) anti-α4 integrin antibodies on oxazolone-induced DC accumulation in lymph nodes in vivo. Antibody was administered to groups of 10 mice in single 100 μl injections intraperitoneally for the 10, 20, and 200 μg doses or in single 30 μl injections intradermally into the dorsum of both ears for 12 μg dose. The 200 μg dose of anti-α4 integrin antibody was given in 2 × 100 μg doses 2 h before and 8 h after oxazolone treatment. Each graph represents a separate experiment and the amount of antibody received per mouse, and the method of administration is indicated below each graph. 2 h after antibody administration, mice received 25 μl of 0.5% oxazolone on the dorsum of both ears. Draining auricular lymph nodes were excised 18 h later and the number of DCs per lymph node was determined. Results for isotype-matched control antibody (hatched bars) and integrin antibody (solid bars) are compared with untreated, naive mice (open bars).

Mentions: If α6 integrins are required for LC migration from the epidermis, in vivo administration of anti-α6 integrin antibody should also inhibit the accumulation of DCs in draining lymph nodes. Anti-α6 integrin antibody was administered either systemically, or directly into the dermis of ear skin, 2 h before topical application of oxazolone. 18 h after exposure to oxazolone, draining auricular lymph nodes were removed and the number of DCs per node was determined. The results from three separate experiments are presented in Fig. 7. In individual experiments, the number of DCs in untreated, naive mice ranged from 2,000 to 5,000/ node. Topical application of oxazolone consistently increased the number of DCs per lymph node by three- to fourfold. Administration of the anti-α6 antibody, GoH3, significantly reduced the number of DCs per lymph node. For example, oxazolone increased the number of DCs per node from 1,675 to 8,275 in mice treated with control antibody, and intraperitoneal injection of 20 μg of GoH3 reduced DC accumulation to 4,353 per node, representing a 59% inhibition of DC accumulation. 10 μg of GoH3 gave slightly less inhibition at 37% and the higher dose of 40 μg did not further inhibit DC accumulation (data not shown). However, 12 μg of antibody administered intradermally was more effective, inhibiting DC accumulation by 74% (Fig. 7).


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

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

The effect of (A) anti-α6  and (B) anti-α4 integrin antibodies on  oxazolone-induced DC accumulation in lymph nodes in vivo. Antibody was administered to groups of  10 mice in single 100 μl injections  intraperitoneally for the 10, 20, and  200 μg doses or in single 30 μl injections intradermally into the dorsum  of both ears for 12 μg dose. The 200  μg dose of anti-α4 integrin antibody  was given in 2 × 100 μg doses 2 h  before and 8 h after oxazolone treatment. Each graph represents a separate experiment and the amount of antibody received per mouse, and the method of administration is indicated below  each graph. 2 h after antibody administration, mice received 25 μl of 0.5% oxazolone on the dorsum of both ears. Draining auricular lymph nodes were  excised 18 h later and the number of DCs per lymph node was determined. Results for isotype-matched control antibody (hatched bars) and integrin antibody (solid bars) are compared with untreated, naive mice (open bars).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199129&req=5

Figure 7: The effect of (A) anti-α6 and (B) anti-α4 integrin antibodies on oxazolone-induced DC accumulation in lymph nodes in vivo. Antibody was administered to groups of 10 mice in single 100 μl injections intraperitoneally for the 10, 20, and 200 μg doses or in single 30 μl injections intradermally into the dorsum of both ears for 12 μg dose. The 200 μg dose of anti-α4 integrin antibody was given in 2 × 100 μg doses 2 h before and 8 h after oxazolone treatment. Each graph represents a separate experiment and the amount of antibody received per mouse, and the method of administration is indicated below each graph. 2 h after antibody administration, mice received 25 μl of 0.5% oxazolone on the dorsum of both ears. Draining auricular lymph nodes were excised 18 h later and the number of DCs per lymph node was determined. Results for isotype-matched control antibody (hatched bars) and integrin antibody (solid bars) are compared with untreated, naive mice (open bars).
Mentions: If α6 integrins are required for LC migration from the epidermis, in vivo administration of anti-α6 integrin antibody should also inhibit the accumulation of DCs in draining lymph nodes. Anti-α6 integrin antibody was administered either systemically, or directly into the dermis of ear skin, 2 h before topical application of oxazolone. 18 h after exposure to oxazolone, draining auricular lymph nodes were removed and the number of DCs per node was determined. The results from three separate experiments are presented in Fig. 7. In individual experiments, the number of DCs in untreated, naive mice ranged from 2,000 to 5,000/ node. Topical application of oxazolone consistently increased the number of DCs per lymph node by three- to fourfold. Administration of the anti-α6 antibody, GoH3, significantly reduced the number of DCs per lymph node. For example, oxazolone increased the number of DCs per node from 1,675 to 8,275 in mice treated with control antibody, and intraperitoneal injection of 20 μg of GoH3 reduced DC accumulation to 4,353 per node, representing a 59% inhibition of DC accumulation. 10 μg of GoH3 gave slightly less inhibition at 37% and the higher dose of 40 μg did not further inhibit DC accumulation (data not shown). However, 12 μg of antibody administered intradermally was more effective, inhibiting DC accumulation by 74% (Fig. 7).

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