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A predominant IgG4 subclass may be responsible for false-negative direct immunofluorescence in bullous pemphigoid

Background: Bullous pemphigoid (BP) is an immune‐mediated blistering disease, usually characterized immunopathologically by the linear deposition of IgG and C3 along the basement membrane zone (BMZ) of skin. However, positive deposition of C3 but negative staining for IgG on direct immunofluorescenc...

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Bibliographic Details
Published in:Journal of cutaneous pathology 2002-05, Vol.29 (5), p.282-286
Main Authors: Buschman, Kerry E., Seraly, Mark, Thong, H. Y., Deng, Jau-Shyong, Draviam, Rose P., Abernethy, John L.
Format: Article
Language:English
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Summary:Background: Bullous pemphigoid (BP) is an immune‐mediated blistering disease, usually characterized immunopathologically by the linear deposition of IgG and C3 along the basement membrane zone (BMZ) of skin. However, positive deposition of C3 but negative staining for IgG on direct immunofluorescence (DIF) studies has been noted in some patients. Methods: Twelve patients known to have BP but with absence of staining for IgG were included in this study. Frozen sections of skin specimens from the 12 patients were subjected to IgG DIF, as well as a sandwich double antibody method of staining for IgG, IgG subclasses, and light chains. Enzyme‐linked immunosorbent assay (ELISA) using commercially available human IgG subclasses was used to analyze the subclass restriction of FITC‐labeled antihuman IgG conjugates. Results:  Of the 12 skin specimens with positive C3 and negative IgG on DIF, nine were positive for IgG with the double antibody sandwich method. In addition, all 12 specimens had positive linear staining for the subclass IgG4 along the BMZ with this method. There was no IgG light chain restriction. Two commercially obtained antihuman IgG conjugates, both commonly used in our laboratory for DIF testing, were analyzed for separate IgG subclass specificity by ELISA. Both conjugates showed high reactivity to IgG1 and IgG3 with less reactivity to IgG2 and IgG4. Conclusion:  These results suggest that the following factors contribute to false‐negative staining for IgG on DIF in some BP patients: (i): subthreshold IgG in skin specimens; (ii) limited reactivity of commercial antihuman IgG conjugates to the IgG4 subclass; and (iii) decreased sensitivity of DIF compared with double antibody methods for the detection of IgG. The use of sandwich double antibody immunofluorescence methods to test for IgG and/or IgG subclasses may be helpful in definitively diagnosing BP in patients with negative IgG and positive C3 staining on DIF.
ISSN:0303-6987
1600-0560
DOI:10.1034/j.1600-0560.2002.290504.x