RESUMO
Dapsone is frequently effective in cutaneous diseases characterized by antibody deposition and accumulation of neutrophils. We hypothesized that this mechanism of action of dapsone may involve the inhibition of neutrophil adherence to antibody. The neutrophil adherence assay, which measures the binding of neutrophils to basement membrane zone-bound antibody on skin sections, was used to evaluate the effect of dapsone on neutrophil adherence to immunoglobulin A and immunoglobulin G. We evaluated the effect of dapsone on adherence of normal neutrophils to immunoglobulin A and immunoglobulin G from sera of linear immunoglobulin A bullous dermatosis and bullous pemphigoid patients, respectively. Linear immunoglobulin A bullous dermatosis or bullous pemphigoid antibody were bound to the basement membrane zone of normal skin sections as a substrate for the neutrophil adherence assay. Dapsone was added directly to the neutrophils or to the antibody source in concentrations of 0-50 micrograms/ml (pharmacologic range). Addition of dapsone to neutrophils produced an incremental inhibition of neutrophil adherence up to 75% at 50 micrograms/ml. Dapsone produced similar inhibition when added directly to the antibody itself, despite washing prior to usage in the neutrophil-adherence assay. Control specimens including irrelevant fractions of patient sera failed to demonstrate binding. Serum from a patient on dapsone therapy also showed inhibition of neutrophil adherence compared to the same patient on no therapy. We conclude that dapsone inhibits the adherence of neutrophils to basement membrane zone antibody in a dose-dependent manner. This may be related to an effect directly on antibody. This inhibition may contribute to the clinical efficacy of dapsone in antibody-mediated diseases.
Assuntos
Dapsona/farmacologia , Imunoglobulina A/análise , Neutrófilos/citologia , Dermatopatias Vesiculobolhosas/imunologia , Adulto , Membrana Basal/imunologia , Membrana Basal/metabolismo , Sítios de Ligação de Anticorpos/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Imunoglobulina A/efeitos dos fármacos , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Penfigoide Bolhoso/imunologia , Dermatopatias Vesiculobolhosas/tratamento farmacológicoRESUMO
Although two groups of bullous pemphigoid antigens have been well characterized, different research groups have shown strikingly different prevalence rates of antibodies to these antigens in their patients. Potential explanations for this phenomena include a patient population that has different prevalence of antibodies, or that the antigen preparations used by the different groups contain different relative amounts of these antigens. We have compared the relative concentration of the different bullous pemphigoid antigens in epidermal extract preparations made by three different procedures commonly used to separate dermis from epidermis: NaCl, ethylenediaminetetraacetic acid (EDTA), and heat. We have found that the amount of the 180-kD antigen present in extracts is dependent on the techniques involved in separation of the epidermis from dermis. NaCl- and EDTA-separation procedures result in partial proteolysis of the 180-kD antigen to smaller forms, including major brands at 160 kD and 97 kD in the EDTA preparation. Fragments of the 180-kD antigen are present in both the separation and wash fluids, associated with a significant reduction of the 180-kD form in the extract of the NaCl-separated skin. We conclude that the native molecular weight of the previously described minor bullous pemphigoid antigen is 180 kD, and that the apparent difference in patient reaction to the 180-kD antigen may be due to different preparations of the antigen rather than underlying differences in seropositivity in the patient population.