| Budget Amount *help |
¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
Type VII collagen is one of the major components of the epidermal basement membrane structure, and is the pathogenic autoantigen of epidermolysis bullosa acquisita (EBA). Precise pathomechanisims in EBA still remains unclear because the reliable model animals for EBA have not been established. The aim of this study is to establish an animal model for EBA, and analyze the pathomechanisms in EBA using the animal model.
First, type VII collagen humanized mice, which express human type VII collagen, but not mouse type VII collagen, in the basement membrane, were immunized with recombinant mouse type VII collagen. However, the anti mouse type VII antibody was not stably produced. In this immunization protocol Then, type VII collagen humanized mice were immunized by grafting the skin from C57BL/6 mice. Indirect immunofluorescence study demonstrated anti mouse types collagen antibody production at 3 weeks after the skin grafting. Major subclasses of the anti mouse type VII collagen antibody were mouse IgG1 and IgG2c. Neither IgG2a nor IgG3 subclass antibodies were present, although the IgG2b subclass antibody was faintly positive.
Next, splenocytes were prepared from anti mouse type VII collagen antibody producing mice, and passively transferred to the immunodeficient RAG2 knockout (KO) mice expressing mouse type VII collagen. Anti mouse type VII collagen antibody was detected 10 days after the transfer, and the recipient mice developed periorbital and perioral erythema and erosions. Histopathological analysis of the lesional skin demonstrated subepidermal blistering and inflammatory cell infiltration at the upper dermis. Direct immunofluorescence analysis revealed the deposition of IgG and C3 at the epidermal basement membrane
This animal model may be useful not only for the further study of EBA pathomechanisms, but also for the development of the novel therapeutic strategy
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