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Osteopontin is involved in the initiation of cutaneous contact hypersensitivity by inducing Langerhans and dendritic cell migration to lymph nodes.

Weiss JM, Renkl AC, Maier CS, Kimmig M, Liaw L, Ahrens T, Kon S, Maeda M, Hotta H, Uede T, Simon JC - J. Exp. Med. (2001)

Bottom Line: Characterizing OPN function for LC/DC migration we found upregulated OPN expression in hapten sensitized skin and draining lymph nodes.OPN induces chemotactic LC/DC migration, initiates their emigration from the epidermis, and attracts LCs/DCs to draining lymph nodes by interacting with CD44 and alphav integrin.In conclusion, OPN is an important factor in the initiation of CHS by guiding LCs/DCs from skin into lymphatic organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Freiburg, D-79104 Freiburg, Germany. weiss@haut.ukl.uni-freiburg.de

ABSTRACT
Osteopontin (OPN) is a chemotactic protein that attracts immune cells, to inflammatory sites. The sensitization phase of allergic cutaneous contact hypersensitivity (CHS) requires the migration of Langerhans cells/dendritic cells (LCs/DCs) from skin to draining lymph nodes. Characterizing OPN function for LC/DC migration we found upregulated OPN expression in hapten sensitized skin and draining lymph nodes. OPN induces chemotactic LC/DC migration, initiates their emigration from the epidermis, and attracts LCs/DCs to draining lymph nodes by interacting with CD44 and alphav integrin. Furthermore, OPN-deficient mice have a significantly reduced CHS response that correlates with an impaired ability of OPN-deficient mice to attract LCs/DCs to draining lymph nodes. In conclusion, OPN is an important factor in the initiation of CHS by guiding LCs/DCs from skin into lymphatic organs.

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OPN is upregulated in skin and lymph nodes during CHS TNCB or acetone was applied to abdominal skin of C57BL/6 mice. Mice were killed at the indicated time points and skin specimens (A) from the haptenized site and axillary lymph nodes draining this skin site (B) or distant maxillary (C) lymph nodes were collected and subjected to RT-PCR with OPN specific primers as described in Materials and Methods. Percent relative intensity of the OPN mRNA compared with β-actin control was calculated with E.A.S.Y.® Win32-software. (D) TNCB was applied to abdominal skin of C57BL/6 mice. Immunohistochemical staining of cryosections with an antibody against OPN or CD3 of skin draining axillary lymph nodes from OPN deficient (−/−) and wild-type (+/+) C57BL/6 mice at the indicated time points after TNCB application to the abdominal skin. Top row: upregulation of OPN expression from 0 to 48 h. Bottom row left: higher magnification of OPN expression at 48 h. Bottom row middle: staining of skin draining axillary lymph nodes from OPN-deficient (−/−) mouse with mAb against OPN. Bottom row right: staining with an antibody against CD3 of a serial section of the lymph node specimen taken at 48 h after TNCB application (top row right), * T cell areas, ** B cell follicle. Magnification, bar 60 μm, except bottom left (48 h) bar 20 μm. OPN was visualized with brown, CD3 with blue chromogen as described in Materials and Methods.
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fig2: OPN is upregulated in skin and lymph nodes during CHS TNCB or acetone was applied to abdominal skin of C57BL/6 mice. Mice were killed at the indicated time points and skin specimens (A) from the haptenized site and axillary lymph nodes draining this skin site (B) or distant maxillary (C) lymph nodes were collected and subjected to RT-PCR with OPN specific primers as described in Materials and Methods. Percent relative intensity of the OPN mRNA compared with β-actin control was calculated with E.A.S.Y.® Win32-software. (D) TNCB was applied to abdominal skin of C57BL/6 mice. Immunohistochemical staining of cryosections with an antibody against OPN or CD3 of skin draining axillary lymph nodes from OPN deficient (−/−) and wild-type (+/+) C57BL/6 mice at the indicated time points after TNCB application to the abdominal skin. Top row: upregulation of OPN expression from 0 to 48 h. Bottom row left: higher magnification of OPN expression at 48 h. Bottom row middle: staining of skin draining axillary lymph nodes from OPN-deficient (−/−) mouse with mAb against OPN. Bottom row right: staining with an antibody against CD3 of a serial section of the lymph node specimen taken at 48 h after TNCB application (top row right), * T cell areas, ** B cell follicle. Magnification, bar 60 μm, except bottom left (48 h) bar 20 μm. OPN was visualized with brown, CD3 with blue chromogen as described in Materials and Methods.

Mentions: Our explant model mimics the migration process of LCs during the sensitization phase of allergic CHS. We therefore speculated that OPN is upregulated in skin during the sensitization phase of CHS. To test this hypothesis, the hapten TNCB was applied to the abdominal skin of C57BL/6 mice. Hapten painted abdominal skin, the regional skin draining axillary lymph nodes, and maxillary lymph nodes distant to the application site (as control) were analyzed for OPN mRNA expression using RT-PCR after 0, 6, 12, 24, and 48 h of TNCB application (Fig. 2, A–C) . 12 h after TNCB painting, OPN mRNA was gradually upregulated in hapten treated skin (Fig. 2 A), peaking at 48 h, in contrast to skin treated with the vehicle acetone alone. In parallel, OPN mRNA was strongly upregulated in axillary lymph nodes draining the skin (Fig. 2 B). In contrast, maxillary lymph nodes distant to the site of hapten application showed only a slight upregulation of OPN mRNA (Fig. 2 C). When immunohistochemistry of skin draining lymph nodes was performed with mAbs against OPN (mAb 2.2 [37]), lymph nodes of untreated (Fig. 2 D) or acetone painted mice (data not shown) showed little to no OPN specific staining. After 24 h following TNCB application, OPN staining was observed in draining lymph nodes which strongly increased up to 48 h (Fig. 2 D). OPN expression was predominantly located in the T cell areas of these lymph nodes as detected by staining of serial lymph node sections with anti-CD3 mAb (Fig. 2 D). No staining was found in lymph nodes of hapten-treated OPN-deficient mice (Fig. 2 D). These findings argue for an OPN function in the sensitization phase of CHS.


Osteopontin is involved in the initiation of cutaneous contact hypersensitivity by inducing Langerhans and dendritic cell migration to lymph nodes.

Weiss JM, Renkl AC, Maier CS, Kimmig M, Liaw L, Ahrens T, Kon S, Maeda M, Hotta H, Uede T, Simon JC - J. Exp. Med. (2001)

OPN is upregulated in skin and lymph nodes during CHS TNCB or acetone was applied to abdominal skin of C57BL/6 mice. Mice were killed at the indicated time points and skin specimens (A) from the haptenized site and axillary lymph nodes draining this skin site (B) or distant maxillary (C) lymph nodes were collected and subjected to RT-PCR with OPN specific primers as described in Materials and Methods. Percent relative intensity of the OPN mRNA compared with β-actin control was calculated with E.A.S.Y.® Win32-software. (D) TNCB was applied to abdominal skin of C57BL/6 mice. Immunohistochemical staining of cryosections with an antibody against OPN or CD3 of skin draining axillary lymph nodes from OPN deficient (−/−) and wild-type (+/+) C57BL/6 mice at the indicated time points after TNCB application to the abdominal skin. Top row: upregulation of OPN expression from 0 to 48 h. Bottom row left: higher magnification of OPN expression at 48 h. Bottom row middle: staining of skin draining axillary lymph nodes from OPN-deficient (−/−) mouse with mAb against OPN. Bottom row right: staining with an antibody against CD3 of a serial section of the lymph node specimen taken at 48 h after TNCB application (top row right), * T cell areas, ** B cell follicle. Magnification, bar 60 μm, except bottom left (48 h) bar 20 μm. OPN was visualized with brown, CD3 with blue chromogen as described in Materials and Methods.
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Related In: Results  -  Collection

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fig2: OPN is upregulated in skin and lymph nodes during CHS TNCB or acetone was applied to abdominal skin of C57BL/6 mice. Mice were killed at the indicated time points and skin specimens (A) from the haptenized site and axillary lymph nodes draining this skin site (B) or distant maxillary (C) lymph nodes were collected and subjected to RT-PCR with OPN specific primers as described in Materials and Methods. Percent relative intensity of the OPN mRNA compared with β-actin control was calculated with E.A.S.Y.® Win32-software. (D) TNCB was applied to abdominal skin of C57BL/6 mice. Immunohistochemical staining of cryosections with an antibody against OPN or CD3 of skin draining axillary lymph nodes from OPN deficient (−/−) and wild-type (+/+) C57BL/6 mice at the indicated time points after TNCB application to the abdominal skin. Top row: upregulation of OPN expression from 0 to 48 h. Bottom row left: higher magnification of OPN expression at 48 h. Bottom row middle: staining of skin draining axillary lymph nodes from OPN-deficient (−/−) mouse with mAb against OPN. Bottom row right: staining with an antibody against CD3 of a serial section of the lymph node specimen taken at 48 h after TNCB application (top row right), * T cell areas, ** B cell follicle. Magnification, bar 60 μm, except bottom left (48 h) bar 20 μm. OPN was visualized with brown, CD3 with blue chromogen as described in Materials and Methods.
Mentions: Our explant model mimics the migration process of LCs during the sensitization phase of allergic CHS. We therefore speculated that OPN is upregulated in skin during the sensitization phase of CHS. To test this hypothesis, the hapten TNCB was applied to the abdominal skin of C57BL/6 mice. Hapten painted abdominal skin, the regional skin draining axillary lymph nodes, and maxillary lymph nodes distant to the application site (as control) were analyzed for OPN mRNA expression using RT-PCR after 0, 6, 12, 24, and 48 h of TNCB application (Fig. 2, A–C) . 12 h after TNCB painting, OPN mRNA was gradually upregulated in hapten treated skin (Fig. 2 A), peaking at 48 h, in contrast to skin treated with the vehicle acetone alone. In parallel, OPN mRNA was strongly upregulated in axillary lymph nodes draining the skin (Fig. 2 B). In contrast, maxillary lymph nodes distant to the site of hapten application showed only a slight upregulation of OPN mRNA (Fig. 2 C). When immunohistochemistry of skin draining lymph nodes was performed with mAbs against OPN (mAb 2.2 [37]), lymph nodes of untreated (Fig. 2 D) or acetone painted mice (data not shown) showed little to no OPN specific staining. After 24 h following TNCB application, OPN staining was observed in draining lymph nodes which strongly increased up to 48 h (Fig. 2 D). OPN expression was predominantly located in the T cell areas of these lymph nodes as detected by staining of serial lymph node sections with anti-CD3 mAb (Fig. 2 D). No staining was found in lymph nodes of hapten-treated OPN-deficient mice (Fig. 2 D). These findings argue for an OPN function in the sensitization phase of CHS.

Bottom Line: Characterizing OPN function for LC/DC migration we found upregulated OPN expression in hapten sensitized skin and draining lymph nodes.OPN induces chemotactic LC/DC migration, initiates their emigration from the epidermis, and attracts LCs/DCs to draining lymph nodes by interacting with CD44 and alphav integrin.In conclusion, OPN is an important factor in the initiation of CHS by guiding LCs/DCs from skin into lymphatic organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of Freiburg, D-79104 Freiburg, Germany. weiss@haut.ukl.uni-freiburg.de

ABSTRACT
Osteopontin (OPN) is a chemotactic protein that attracts immune cells, to inflammatory sites. The sensitization phase of allergic cutaneous contact hypersensitivity (CHS) requires the migration of Langerhans cells/dendritic cells (LCs/DCs) from skin to draining lymph nodes. Characterizing OPN function for LC/DC migration we found upregulated OPN expression in hapten sensitized skin and draining lymph nodes. OPN induces chemotactic LC/DC migration, initiates their emigration from the epidermis, and attracts LCs/DCs to draining lymph nodes by interacting with CD44 and alphav integrin. Furthermore, OPN-deficient mice have a significantly reduced CHS response that correlates with an impaired ability of OPN-deficient mice to attract LCs/DCs to draining lymph nodes. In conclusion, OPN is an important factor in the initiation of CHS by guiding LCs/DCs from skin into lymphatic organs.

Show MeSH
Related in: MedlinePlus