Indirect complement immunofluorescence in the immunopathological assessment of bullous pemphigoid, cicatricial pemphigoid, and herpes gestationis

Abstract

Summary Sera from 50 untreated patients with bullous pemphigoid were examined by both the customary method of indirect immunofluorescence using anti IgG conjugate and by indirect complement immunofluorescence using anti C3 conjugate to detect circulating antibasement membrane zone antibodies. A circulating antibasement membrane zone antibody could be detected by the IgG method in 58% and by the C3 method in 76%. Sera from six patients with cicatricial pemphigoid examined in the same way showed a circulating antibasement membrane zone antibody in one by the IgG method but in three by the C3 method of indirect immunofluorescence. Sera from ten patients with active herpes gestationis contained anti‐basement membrane zone antibody demonstrable by the C3 method in every case and by the IgG method of indirect immunofluorescence in one of these. Basement membrane zone bound IgG, or more commonly C3 in a linear pattern, was shown by direct immunofluorescence in all patients with bullous or cicatricial pemphigoid from whom adequate biopsy material was obtained. The immunopathological similarities of bullous pemphigoid, cicatricial pemphigoid and herpes gestationis are stressed, and the usefulness of indirect complement immunofluorescence in their diagnosis is emphasized.

Clinical and immunological profiles of 25 patients with pemphigoid gestationis

“…18 We also performed complement IF of both normal and 1 mol L À1 NaCl-split skin as described previously. 19,20 Immunoblotting (IB) analyses of normal human epidermal extracts, the recombinant proteins (RPs) of the NC16a and C-terminal domains of BP180, and concentrated culture supernatant of HaCaT cells were performed as previously described. 8,20,21 Enzyme-linked immunosorbent assays (ELISAs) of the NC16a domain of BP180 RP, and of the N-and C-terminal domains of BP230 RP were performed using commercially available kits (MESACUP; MBL, Nagoya, Japan).…”

Bullous pemphigoid: the frequency of mucosal involvement and concurrent malignancy related to indirect immunofluorescence findings

“…Immunofluorescence techniques for the demonstration of anti-BMZ antibodies followed standard procedures and have been described elsewhere (Beutner et al, 1973;Hodge et al, 1978). Briefiy, serum was tested for the presence of an anti-BMZ antibody in dilutions of not less than i: 10 using commercially obtained anti-IgG conjugate against human skin substrate.…”

Sequential Intramolecular Epitope Spreading of Humoral Responses to Human BPAG2 in a Transgenic Model

“…Serial dilutions of murine sera were tested against cryosections of intact or 1 M NaCl split human skin by indirect and complement-binding IF microscopy as described previously (Hodge et al, 1978). Fluorescein-conjugated secondary antibodies diluted 1:20 (rat anti-mouse IgE (g chain specific) and goat anti-mouse IgA (a chain specific) (SouthernBiotech, Chicago, IL)) were used.…”