Immunologic and Histopathologic Characterization of an Active Disease Mouse Model for Pemphigus Vulgaris Manabu Ohyama, Masayuki Amagai, Kazuyuki Tsunoda,

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Immunologic and Histopathologic Characterization of an Active Disease Mouse Model for Pemphigus Vulgaris Manabu Ohyama, Masayuki Amagai, Kazuyuki Tsunoda, Takayuki Ota, Shigeo Koyasu, Jun-ichi Hata, Akihiro Umezawa, Takeji Nishikawa Journal of Investigative Dermatology Volume 118, Issue 1, Pages 199-204 (January 2002) DOI: 10.1046/j.0022-202x.2001.01643.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Predominant IgG1 deposition on keratinocyte surfaces in PV model mice in vivo. (a) Strong IgG and very weak IgA deposition was observed on the surfaces of keratinocytes of the palate of PV model mice. F, fibrinogen. (b) IgG1 was predominantly observed in the intercellular space. IgG2a and IgG2b were weakly detected and no apparent IgG3 deposition was observed. Scale bar: 50 µm. Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Circulating IgG1 against mouse Dsg3 predominates in sera obtained from PV model mice. (a) All 31 PV model mice sera were positive for anti-Dsg3 IgG, whereas all were negative for anti-Dsg1 IgG. Three and one sera were slightly positive for anti-Dsg3 IgA and IgM, respectively. (b) All 20 PV model mice sera showed positive in IgG1 ELISA, whereas 12 sera were positive for IgG2a and IgG2b. No sera were positive for IgG3. Bars indicate the cut-off values determined for individual ELISA. Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Gross phenotypes of PV model and Dsg3–/– mice. Compared with the wild-type B6 mouse (a, right), PV model mice (a, left) showed patchy telogen hair loss and weight loss. Dsg3–/– mice also showed patchy telogen hair loss and weight loss (b), similar in extent to the PV model mice. Some PV model mice showed crust-covered severe erosions on the paws, and elongated front teeth (c), whereas Dsg3–/– mice rarely showed these changes (d). Scale bar: 2 cm. Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Histopathologic comparisons of PV model and Dsg3–/– mice. Histopathologic findings in PV model mice (a, c, e, g, i, j) and Dsg3–/– mice (b, d, f, h). Suprabasilar acantholysis was observed with similar distribution on the traumatized skin (a, b, snout; c, d, ear; e, f, paw) and mucous membranes (g, h: hard palate); however, involvement of the esophagus and forestomach was observed only in PV model mice (i, j). Mucosal epithelia were detached from the lamina propria mucosae with inflammatory cell infiltrations (i, esophagus). Acantholysis was observed around the squamocolumnar junction (j, forestomach). Scale bar: 50 µm (hematoxylin and eosin staining). Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Affected sites in PV model and Dsg3–/– mice. The number of affected sites per mouse was counted for PV model and Dsg3–/– mice. Some PV model mice were more severely affected and PV model mice showed broader variations in the extent of microscopic lesions than Dsg3–/– mice. Bars indicate the mean values for each group. Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Eosinophilic spongiosis in PV model mice. Eosinophilic cell infiltrations in the intercellular space of the epidermis were found in some spongiotic sites of PV model mice (a), but not in those of Dsg3–/– mice (b). Arrows indicate eosinophils. Scale bar: 50 µm (Chromotrope 2R and hematoxylin staining). Journal of Investigative Dermatology 2002 118, 199-204DOI: (10.1046/j.0022-202x.2001.01643.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions