Immunoblotting(IB) technique has been frequently used to detect bullous pemphigoid (BP) antibodies in BP patient.It is important to extract the antigen proteins fully and intactly in the base membrane zone(BMZ) of normal human epidermis.The simple method of extracting antigens will make IB techniqne easy.Three different methods of extracting antigen proteins were compared.Using IB for detecting BP antibodies,it has been found that the results of epidermal extract preparations are basically similar.However,one of these methods (NO.1) is more convenient and may obtain more amounts of antigens.Another method(No.3) is identical to that of extracting antigens in dermis for de- tecting epidermolysis bullosa acquista (EBA) antibodies.Therefore,in using IB technique to distinguish BP from EBA,one can apply method No.3.

Bullous pemphigoid (BP) antigens are of molecular heterogeneity. To find out the relationship between the antigenic molecular heterogeneity and clinical features, treatment and prognosis in BP, 122 sera of BP patients were tested with immunoblotting technique. The results showed that 94 sera reacted with BP antigens extracted from normal human epidermis. Among them, 54 sera (57.4%) had BP 230kD antibody and 83 sera (88.3%) had 180kD antibody. Based on the results of immunoblotting assay, these 94 BP patients who had the positive antibodies were divided into three groups, including group I patients with 230kD protein antibody only, group Ⅱ patients with 180kD protein antibody and other minor protein antibodies, and group Ⅲ patients with both 230kD and 180kD antibodies. The pattern of clinical presentations (mucosal involvement, scarring and milia), disease severity, the largest dose of prednisone per day and duration of hospitalization were compared in these three groups. The results of comparison showed that there was no statistically significant difference among three groups in above mentioned items, except the difference of the largest dose of prednisone between the group Ⅰ and group Ⅱ (t=2.22, P

In order to investigate the relationship between lichen planus pemphigoides (LPP) and bullous pemphigoid (BP) and to determine whether the LPP antigen is the classical BP antigen. Four LP patients who were found to have blisters or subepidermal bullae by clinical and histopathological findings were investigated using DIF, IIF, immunoblotting methods. The results showed that there were linear deposition of IgG along the BMZ in three cases, C3 granular deposition in four cases, and linear fibrin deposition in 3 cases. Two out of these patients showed circulating IgG anti-BMZ antibody with titers 1:80 and 1:320, and lmol/L NaCl split section showed that the antibody was bound to the epidermal side, as did control BP antisera. Western blotting of the epidermal extracts using the sera of these 4 patients showed that all 4 patients'sera reacted with the epidermal antigen, and the sera of 2 patients reacted with 200kD antigen which is different from classical bullous pemphigoid antigen. With a review of literature we suggest that LPP is not the overlapping of BP and LP.