IgG anti-laminin-332 autoantibodies are present in a subset of patients with mucous membrane, but not bullous, pemphigoid Zelmira Lazarova, MD, Valerie.

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IgG anti-laminin-332 autoantibodies are present in a subset of patients with mucous membrane, but not bullous, pemphigoid Zelmira Lazarova, MD, Valerie K. Salato, MS, Christoph M. Lanschuetzer, MD, Marleen Janson, BS, Janet A. Fairley, MD, Kim B. Yancey, MD Journal of the American Academy of Dermatology Volume 58, Issue 6, Pages 951-958 (June 2008) DOI: 10.1016/j.jaad.2008.02.035 Copyright © 2008 American Academy of Dermatology, Inc. Terms and Conditions

Fig 1 IgG4 anti-laminin-332 enzyme-linked immunosorbent assay (ELISA) detects autoantibodies in patients with antiepiligrin cicatricial pemphigoid (AECP) with great sensitivity and specificity. Scatter plot representation of ELISA results concerning performance of sera from 32 patients with AECP and 173 control subjects. All sera were tested in duplicate; plotted dots represent average of OD490 reading obtained for each sample. Dashed line indicates cut-point score determined by receiver operating characteristic (ROC) methodology. Insert depicts ROC curve of assay (sensitivity 0.91 [confidence interval {CI} 0.75-0.98], specificity 0.98 [CI 0.95-0.99], Youden index 0.89). BP, Bullous pemphigoid; NHS, normal human serum; PF, pemphigus foliaceus; PV, pemphigus vulgaris. Journal of the American Academy of Dermatology 2008 58, 951-958DOI: (10.1016/j.jaad.2008.02.035) Copyright © 2008 American Academy of Dermatology, Inc. Terms and Conditions

Fig 2 Sera from patients with bullous pemphigoid (BP) do not contain IgG autoantibodies directed against laminin (L)-332. A, Sera from 13 patients with BP scoring above cut point in IgG4 anti-L-332 enzyme-linked immunosorbent assay (ELISA) (lanes 1-13) immunoblotted either BP antigen (BPAG)1 or BPAG2 in human keratinocyte (HK) extracts. None of these samples displayed reactivity against L-332. Lane L represents positive control where L-332 was immunoblotted with well-characterized rabbit antiserum and developed with alkaline phosphatase-conjugated goat antirabbit IgG; lane N represents negative control where L-332 was immunoblotted with normal human serum and developed with same second-step antibody used in immunoblot studies of sera from patients with BP. Ticks in left margin correspond to BPAG1 and BPAG2; brackets in right margin correspond to various L-332 subunits (specifically, unprocessed and processed α subunit, unprocessed and processed γ subunit, and β subunit). B, Sera from 13 patients with BP scoring above cut point in IgG4 anti-L-332 ELISA (lanes 1-13) showed no evidence of reactivity to L-332 in HK extracellular matrix (most sensitive antigen source for detection of autoantibodies in patients with antiepiligrin cicatricial pemphigoid [AECP]).15Lanes L and N and brackets in right margin are same as noted in description of A. C, Sera from all patients with BP scoring above cut point in IgG4 anti-L-332 ELISA (lanes 1-13) immunoprecipitated BPAG1, BPAG2, or both from extracts of biosynthetically radiolabeled HKs. None of these samples immunoprecipitated L-332 from this antigen source (most sensitive known technique for detection of autoantibodies in patients with AECP).15Lane L represents positive control where L-332 was immunoprecipitated with well-characterized rabbit antiserum. Annotations in panel margins are same as those depicted in A. Journal of the American Academy of Dermatology 2008 58, 951-958DOI: (10.1016/j.jaad.2008.02.035) Copyright © 2008 American Academy of Dermatology, Inc. Terms and Conditions