Establishment of the mouse model by rabbit antiserum specific to EC1-2 or EC3-4 epitopes for studying the pathogenesis of pemphigus vulgaris.
The research goal was to establish the neonatal mouse model by specific rabbit anti EC1-2 or EC3-4 antiserum for the purpose of studying pemphigus vulgaris (PV) pathogenesis. RNA was extracted from human keratinocytes. The cDNAs was synthesized by reverse transcription. Amplified EC1-2 or EC3-4 genes were inserted into pGEX-4T expression plasmids to constitute the recombinant plasmids, and transformed into E.coli. for the expression of the fusion proteins. The purified fusion proteins were used to immunize New Zealand white rabbits to obtain the specific anti EC1-2 or EC3-4 antisera. IgG fractions from the antisera were purified and passively transferred intradermally into the upper back skin of neonatal BALB/c mice. Histologic, ultrastructural, and immunofluorescence features of PV were evaluated in these mice. PV features were shown in the mice injected with anti EC1-2 antibody, erythema on the flanks and positive Nikolsky sign. The histology showed intraepidermal vesicle formation and acantholytic cells within. Acantholysis, but no clinical symptoms, was seen in the mice treated with the antibody specific to EC3-4. Additionally, skin sections from the abdomen from these neonatal mice were cryo-sectioned for direct-immunofluorescence. FITC-conjugated IgG antibody deposit between the keratinocytes throughout the epidermis. The indirect immunofluorescence with monkey esophagus showed the presence of anti-intracellular antibody with a titer of 1:40. On electronic microscopy, intercellular spaces (ICS) were widened and the desmosomes were split or dissolved. A novel PV mouse model was established by treatment with the specific rabbit antiserum. These data confirmed that both EC1-2 and EC3-4 are pathogenic epitopes in PVA, but EC1-2 is more dominant.
Department of Dermatology, Rui Jin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China, 200025.
Meng
M
Pan
Xiaoqing
X
Zhao
Feng
F
Xue
Weiping
W
Li
Jie
J
Zheng
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015820:Cadherins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D004195:Disease Models, Animal; D007106:Immune Sera / Q000276:immunology / Q000633:toxicity*; D016056:Immunodominant Epitopes / Q000276:immunology*; D051379:Mice / Q000276:immunology*; D008807:Mice, Inbred BALB C; D010392:Pemphigus / Q000276:immunology* / Q000473:pathology; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D011817:Rabbits
D000906:Antibodies; D015820:Cadherins; D007106:Immune Sera; D016056:Immunodominant Epitopes