Pathogenicity of autoantibodies in anti-p200 pemphigoid.
Recently, the C-terminus of laminin γ1 has been identified as target antigen in anti-p200 pemphigoid and the disease was renamed as anti-laminin γ1 pemphigoid. However, the pathogenic relevance of these autoantibodies has not yet been demonstrated. Therefore, we employed an ex vivo model of autoantibody-mediated leukocyte-dependent neutrophil activation and dermal-epidermal separation (DES) using cryosections of human skin. We showed that anti-p200 pemphigoid sera (n = 7) induced DES in a time-dependent manner, in contrast to sera from healthy controls. Furthermore, laminin γ1-specific IgG and serum depleted from anti-laminin γ1 reactivity were generated using the recombinant C-terminus of laminin γ1 (LAMC1-term; amino acids 1364 to 1609). Interestingly, both fractions labeled the dermal-epidermal-junction (DEJ) by indirect immunofluorescence microscopy on human foreskin and recognized a 200 kDa protein by immunoblotting with dermal extract. Human and rabbit IgG against LAMC1-cterm failed to attract neutrophils at the DEJ and to induce DES. In contrast, patient serum depleted from LAMC1-cterm reactivity led to the same extent of DES as non-depleted IgG. Repeated injection of rabbit anti-murine LAMC1-cterm IgG into both neonatal and adult C57BL/6mice as well as repetitive immunization of various mouse strains with murine LAMC1-cterm failed to induce macro- and microscopic lesions. In all mice, circulating anti-LAMC1-cterm antibodies were present, but only in some mice, IgG deposits were seen at the DEJ. We conclude that autoantibodies in anti-p200 pemphigoid sera are pathogenic while pathogenicity is not mediated by autoantibodies against laminin γ1. Further studies are needed to identify the pathogenically relevant autoantigen in anti-p200 pemphigoid.
Department of Dermatology, University of Luebeck, Luebeck, Germany.
Katerina
K
Vafia
Stephanie
S
Groth
Tina
T
Beckmann
Misa
M
Hirose
Jenny
J
Dworschak
Andreas
A
Recke
Ralf J
RJ
Ludwig
Takashi
T
Hashimoto
Detlef
D
Zillikens
Enno
E
Schmidt
D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D001323:Autoantibodies / Q000096:biosynthesis / Q000097:blood / Q000276:immunology*; D001768:Blister / Q000097:blood / Q000276:immunology / Q000473:pathology; D016616:Cryoultramicrotomy; D018076:DNA, Complementary / Q000235:genetics; D020405:Dermis / Q000276:immunology / Q000473:pathology; D004817:Epidermis / Q000276:immunology / Q000473:pathology; D018604:Epitope Mapping; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D007116:Immunization, Passive; D007074:Immunoglobulin G / Q000276:immunology; D007797:Laminin / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010391:Pemphigoid, Bullous / Q000097:blood / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology
D001323:Autoantibodies; D018076:DNA, Complementary; D007106:Immune Sera; D007074:Immunoglobulin G; D007797:Laminin; D011994:Recombinant Proteins; C089141:laminin gamma 1