Molecular and functional characterization of the four-transmembrane molecule l6 in epidermal keratinocytes.
In normal human epidermal keratinocytes (NHEK) proteolytic detachment from the substrate induces a complex activation cascade including expression of new proteins, morphological alterations, and the onset of migration for epidermal regeneration. By subtractive cloning we have shown that L6, a four-transmembrane protein, is newly expressed after proteolytic keratinocyte detachment. In this study, we have generated a novel anti-L6 antibody (clone HD-pKe#104-1.1) and investigated L6 expression regulation in vitro and in vivo as well as L6 function in keratinocyte migration. Dispase-mediated detachment induced L6 expression in NHEK at the mRNA and protein level. Immunohistology of skin biopsies displayed a strong expression of L6 in follicular epidermis and epidermolytic lesions of autoimmune bullous dermatoses (bullous pemphigoid, pemphigus vulgaris), but not in normal interfollicular epidermis. In contrast to normal keratinocytes, HaCaT cells showed constitutive L6 expression, indicating a constitutively active phenotype. After artificial wounding of confluent HaCaT cultures, anti-L6 antibody strongly impaired cell migration velocity and migratory reepithelization of the defect, indicating L6 involvement in keratinocyte migration. These findings suggest that L6 is an important activation-dependent regulator of keratinocyte function and epidermal tissue regeneration.
Storim
J
J
Institute for Immunology, University of Heidelberg, Im Neuenheimer Feld 305, Heidelberg, 69120, Germany.
Friedl
P
P
Schaefer
B M
BM
Bechtel
M
M
Wallich
R
R
Kramer
M D
MD
Reinartz
J
J
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D015603:Keratinocytes / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010391:Pemphigoid, Bullous / Q000473:pathology; D010392:Pemphigus / Q000473:pathology; D011994:Recombinant Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; C074075:TM4SF1 protein, human