Fig 1.
Immunization with murine desmoglein 3 induces antibodies against Dsg3 but no clinical PV phenotype.
(A) C57Bl/6J, B6.SJL-H2s, DBA2/J or SJL/J mice were immunized 4x with mDsg3 emulsified in TiterMax™ on days 0, 14, 35, 56. (B) Dsg3-specific IgG autoantibody titers of C57Bl/6J (n = 4), B6.SJL-H2s (n = 4), DBA2/J (n = 4) and SJL/J (n = 4) mice immunized 4x with mDsg3 emulsified in TiterMax™ on days 0, 14, 35, and 56. (C) Direct immunofluorescent staining for anti-mouse-IgG (facial skin) at day 84. (D) Monkey esophagus was incubated with serum from immunized mice (day 84) and stained for anti-mouse-IgG. In both direct and indirect IF microscopy, no binding of IgG to the epidermis was detected. Dotted line: Dermal-epidermal junction. (E) Representative picture of endoscopy of the oral cavity showed no mucous membrane lesions at day 84. (F) The formation of mDsg3-specific B or plasma cells was confirmed by immunofluorescence of draining lymph nodes (day 84). B220: red, recombinant mDsg3 protein: green, arrows indicate Dsg3-specific B cells.
Fig 2.
Immunization with murine desmoglein 3 and additional ETA or human desmoglein induce antibodies against Dsg3, but no clinical PV phenotype.
(A) Modified 4x immunization protocol (additional injection of 0.5 μg/ml ETA at day 28) was tested in DBA2/J and SJL/J mice on day 0, 14, 35, 56. The s.c. immunization was carried out with 100 μg mDsg3 emulsified in TiterMax™. (B) Dsg3-specific IgG autoantibody titers of DBA2/J mice (n = 4) and SJL/J mice (n = 3). (C) Modified 4x immunization protocol was tested in DBA2/J and SJL/J mice on day 0, 14, 35, 56. The s.c. immunization was carried out with 30 μg human (h) Dsg1 and 70 μg hDsg3 emulsified in TiterMax™ (C). IgG autoantibody titers of DBA2/J mice (n = 4) and SJL/J (n = 5) were analyzed for their activity against (D) hDsg1, (E) hDsg3 and (F) cross-reactivity with mDsg3.