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Diagnosis of anti-laminin γ-1 pemphigoid by immunoblot analysis.
J Eur Acad Dermatol Venereol. 2019 Apr; 33(4):735-741.JE

Abstract

BACKGROUND

Anti-laminin-γ1 (lam-γ1) pemphigoid, a recently described immunobullous disorder sharing immune serological features of bullous pemphigoid and epidermolysis bullosa acquisita (EBA), is characterized by the detection of serum IgG autoantibodies against the lam-γ1 chain, a 200 kDa heterotrimeric component of the dermal-epidermal junction (DEJ).

OBJECTIVE

The aim of the study was to develop an easy-to-perform and reliable assay for the serological detection of anti-lam-γ1 IgG autoantibodies. The clinical appearance alone is not sufficient to establish diagnosis of anti-lam-γ1 pemphigoid and rather requires immune serological evidence of (i) IgG reactivity against the dermal portion of salt-split human skin; (ii) exclusion of IgG against other components of the DEJ; and (iii) IgG reactivity with a 200 kDa protein of dermal extracts by immunoblot analysis (IB).

METHODS

The sera of 55 patients with anti-lam-γ1 pemphigoid were tested by IB with two recombinant heterotrimers, laminin 111 (lam-111) and laminin 421 (lam-421), as well as with a recombinant lam-γ1 chain monomer. Additionally, a total of 41 control sera from patients with EBA (n = 15), psoriasis vulgaris (PV; n = 14), and healthy controls (HC; n = 12) were tested.

RESULTS

Immunoblot analysis revealed a positive reactivity with lam-111 and/or lam-421 in 46/55 (84%) of anti-lam-γ1 pemphigoid sera. Moreover, 8/9 of the initially non-reactive sera were positive with the lam-γ1 monomer, leading to an overall sensitivity of 98.2%. Analyses of 41 control sera with the three lam-γ1 recombinants led to a specificity of 88%. Specifically, 3/15 EBA sera, 1/14 PV serum and 1/12 HC serum reacted with the lam-γ1 monomer while only the 3 EBA sera reacted with lam-421.

CONCLUSIONS

Here we show a novel two-step IB assay using the two recombinant laminin trimers and lam-γ1 chain monomer for the detection of anti-lam-γ1 serum IgG with high sensitivity and specificity. This assay will facilitate the diagnosis and further characterization of this disease.

Authors+Show Affiliations

Department of Dermatology and Allergology, Philipps-University, Marburg, Germany.Department of Dermatology and Allergology, Philipps-University, Marburg, Germany.Kurume University Institute of Cutaneous Cell Biology, Kurume, Fukuoka, Japan.Department of Dermatology and Allergology, Philipps-University, Marburg, Germany.Kurume University Institute of Cutaneous Cell Biology, Kurume, Fukuoka, Japan. Osaka City University Graduate School of Medicine, Asahimachi, Abeno, Osaka, Japan.Department of Dermatology and Allergology, Philipps-University, Marburg, Germany.Department of Dermatology and Allergology, Philipps-University, Marburg, Germany.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29972879

Citation

Solimani, F, et al. "Diagnosis of Anti-laminin Γ-1 Pemphigoid By Immunoblot Analysis." Journal of the European Academy of Dermatology and Venereology : JEADV, vol. 33, no. 4, 2019, pp. 735-741.
Solimani F, Pollmann R, Ishii N, et al. Diagnosis of anti-laminin γ-1 pemphigoid by immunoblot analysis. J Eur Acad Dermatol Venereol. 2019;33(4):735-741.
Solimani, F., Pollmann, R., Ishii, N., Eming, R., Hashimoto, T., Schmidt, T., & Hertl, M. (2019). Diagnosis of anti-laminin γ-1 pemphigoid by immunoblot analysis. Journal of the European Academy of Dermatology and Venereology : JEADV, 33(4), 735-741. https://doi.org/10.1111/jdv.15170
Solimani F, et al. Diagnosis of Anti-laminin Γ-1 Pemphigoid By Immunoblot Analysis. J Eur Acad Dermatol Venereol. 2019;33(4):735-741. PubMed PMID: 29972879.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Diagnosis of anti-laminin γ-1 pemphigoid by immunoblot analysis. AU - Solimani,F, AU - Pollmann,R, AU - Ishii,N, AU - Eming,R, AU - Hashimoto,T, AU - Schmidt,T, AU - Hertl,M, Y1 - 2018/07/30/ PY - 2018/03/12/received PY - 2018/06/01/accepted PY - 2018/7/5/pubmed PY - 2019/8/6/medline PY - 2018/7/5/entrez SP - 735 EP - 741 JF - Journal of the European Academy of Dermatology and Venereology : JEADV JO - J Eur Acad Dermatol Venereol VL - 33 IS - 4 N2 - BACKGROUND: Anti-laminin-γ1 (lam-γ1) pemphigoid, a recently described immunobullous disorder sharing immune serological features of bullous pemphigoid and epidermolysis bullosa acquisita (EBA), is characterized by the detection of serum IgG autoantibodies against the lam-γ1 chain, a 200 kDa heterotrimeric component of the dermal-epidermal junction (DEJ). OBJECTIVE: The aim of the study was to develop an easy-to-perform and reliable assay for the serological detection of anti-lam-γ1 IgG autoantibodies. The clinical appearance alone is not sufficient to establish diagnosis of anti-lam-γ1 pemphigoid and rather requires immune serological evidence of (i) IgG reactivity against the dermal portion of salt-split human skin; (ii) exclusion of IgG against other components of the DEJ; and (iii) IgG reactivity with a 200 kDa protein of dermal extracts by immunoblot analysis (IB). METHODS: The sera of 55 patients with anti-lam-γ1 pemphigoid were tested by IB with two recombinant heterotrimers, laminin 111 (lam-111) and laminin 421 (lam-421), as well as with a recombinant lam-γ1 chain monomer. Additionally, a total of 41 control sera from patients with EBA (n = 15), psoriasis vulgaris (PV; n = 14), and healthy controls (HC; n = 12) were tested. RESULTS: Immunoblot analysis revealed a positive reactivity with lam-111 and/or lam-421 in 46/55 (84%) of anti-lam-γ1 pemphigoid sera. Moreover, 8/9 of the initially non-reactive sera were positive with the lam-γ1 monomer, leading to an overall sensitivity of 98.2%. Analyses of 41 control sera with the three lam-γ1 recombinants led to a specificity of 88%. Specifically, 3/15 EBA sera, 1/14 PV serum and 1/12 HC serum reacted with the lam-γ1 monomer while only the 3 EBA sera reacted with lam-421. CONCLUSIONS: Here we show a novel two-step IB assay using the two recombinant laminin trimers and lam-γ1 chain monomer for the detection of anti-lam-γ1 serum IgG with high sensitivity and specificity. This assay will facilitate the diagnosis and further characterization of this disease. SN - 1468-3083 UR - https://www.unboundmedicine.com/medline/citation/29972879/Diagnosis_of_anti_laminin_γ_1_pemphigoid_by_immunoblot_analysis_ L2 - https://doi.org/10.1111/jdv.15170 DB - PRIME DP - Unbound Medicine ER -