Ultrastructural localization of in vitro binding sites of circulating anti-basement membrane zone antibodies in bullous pemphigoid

Yamasaki, Y.; Nishikawa, T.

doi:10.2340/0001555563501506

Acta Derm Venereol

1983

DOI: 10.2340/0001555563501506

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Ultrastructural localization of in vitro binding sites of circulating anti-basement membrane zone antibodies in bullous pemphigoid

Y. Yamasaki,

T. Nishikawa

Abstract: Ultrastructural localization of the in vitro binding sites of circulating anti-basement membrane zone antibodies was investigated using two immunoperoxidase methods: the ordinary IgG indirect immunoperoxidase method and the complement immunoperoxidase method. The results indicate that the binding sites of the circulating autoantibodies were diverse. It is postulated that those antibodies demonstrable by the ordinary IgG indirect method and those antibodies possessing capacity to fix complement should bind to d… Show more

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Cited by 23 publications

References 16 publications

(25 reference statements)

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1-2B7B: Monoclonal antibody reacting to the 120 kDa polypeptide component of human epidermal hemidesmosomes

et al. 1991

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Immunohistochemical and immunochemical analyses were performed on a monoclonal antibody designated 1-2B7B which was derived from immunizing mice with human prostate epithelial tissue. The 1-2B7B antigen was expressed not only along the acinous basement membrane zone (BMZ) of the prostate and testis, but also along the BMZ of the epithelia of several other organs including the skin, oesophagus, urinary bladder, ureter, stomach, intestine and bile duct. The antigenic epitope was not expressed in these tissues of lower mammals. Immunoelectron microscopic studies on normal human skin revealed that the 1-2B7B antigen was localized mainly just beneath the hemidesmosomes of basal keratinocytes, but not beneath melanocytes. Indirect immunofluorescence and immunoelectron microscopic studies on 1 M NaCl-split skin confirmed that this antigen was not separated from the cytoplasmic membrane of basal cells after salt treatment. SDS polyacrylamide gel electrophoresis of immunochemically purified protein from the epidermis demonstrated the molecular weight of the antigen to be 120 kDa. 1-2B7B monoclonal antibody should be a useful probe for studying the pathomechanism of some blistering diseases, as well as the assembly and function of the epidermal-dermal junction.

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1-2B7B: Monoclonal antibody reacting to the 120 kDa polypeptide component of human epidermal hemidesmosomes

et al. 1991

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Bullous pemphigoid antigen concentration in normal human skin in relation to body area and age

Hamm

Woźniak

1988

Arch Dermatol Res

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The amount of bullous pemphigoid antigen (BPA) in normal human skin on different areas of the body and in bodies of different ages was determined by making endpoint titer estimations using the indirect immunofluorescence technique. Two sera with bullous pemphigoid antibodies were tested with 36 specimens of normal human skin from six cadavers. The greatest expression of BPA was found in plantar sites, whereas the lowest endpoint titers were seen in flexural arm biopsy specimens. Age-dependent differences in BPA expression were detected as well. The oldest individuals tested produced the highest endpoint titers in contrast to the lowest endpoint titers in the youngest. This study suggests that the amount of BPA depends not only on the body region where the biopsy specimen is taken but also on the age of the donor. This is important to know in connection with indirect immunofluorescence assays for BP antibodies on human skin.

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Demonstration of intra- and extracellular localization of bullous pemphigoid antigen using cryofixation and freeze substitution for postembedding immunoelectron microscopy

et al. 1989

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Using low-temperature postembedding techniques for immunoelectron microscopy, we succeeded in demonstrating the precise localization of bullous pemphigoid antigen (BP-Ag) in normal human skin. Small pieces (less than 1 mm3) of normal adult skin were rapidly frozen in liquid propane at -190 degrees C and subjected to freeze substitution with 100% methanol at -80 degrees C. Specimens were embedded in Lowicryl K11M at -60 degrees C which was polymerized under ultraviolet radiation at -60 degrees C. Ultrathin sections were incubated with BP sera followed by rabbit anti-human IgG and colloidal-gold conjugated anti-rabbit IgG. Epidermal ultrastructure was generally well preserved: the basal cell plasma membrane and intra- and extracellular components of hemidesmosomes could be resolved. Gold particles were mainly distributed on and around the hemidesmosomes in both intra- and extracellular sites, with most of the labelling being inside the basal keratinocytes and within about 300 nm of the basal plasma membrane. No specific labelling was observed beneath melanocytes or when normal human serum was used as a control instead of BP serum. Our observations indicate that BP-Ag is localized in and around hemidesmosomes in normal human skin and that the antigen has both intracellular and extracellular domains with the major component occurring inside the cells.

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Ultrastructural localization of in vitro binding sites of circulating anti-basement membrane zone antibodies in bullous pemphigoid

Cited by 23 publications

References 16 publications

1-2B7B: Monoclonal antibody reacting to the 120 kDa polypeptide component of human epidermal hemidesmosomes

1-2B7B: Monoclonal antibody reacting to the 120 kDa polypeptide component of human epidermal hemidesmosomes

Bullous pemphigoid antigen concentration in normal human skin in relation to body area and age

Demonstration of intra- and extracellular localization of bullous pemphigoid antigen using cryofixation and freeze substitution for postembedding immunoelectron microscopy

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