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Mucosal pemphigus vulgaris anti-Dsg3 IgG is pathogenic to the oral mucosa of humanized Dsg3 mice.

Culton DA, McCray SK, Park M, Roberts JC, Li N, Zedek DC, Anhalt GJ, Cowley DO, Liu Z, Diaz LA - J. Invest. Dermatol. (2015)

Bottom Line: There are two major clinical subsets of pemphigus vulgaris (PV)-mucosal PV (mPV) and mucocutaneous PV (mcPV).The majority of mPV sera preferentially recognize hDsg3 compared with mDsg3 by immunoprecipitation as well.These findings suggest that the Dsg3 epitopes targeted by pathogenic mPV IgG are human specific.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

ABSTRACT
There are two major clinical subsets of pemphigus vulgaris (PV)-mucosal PV (mPV) and mucocutaneous PV (mcPV). The mPV subset exhibits anti-human desmoglein (Dsg) 3 autoantibodies that fail to recognize murine Dsg3 (mDsg3); thus, passive transfer experiments of mPV IgG into wild-type (WT) mice have been unsuccessful at inducing disease. We therefore generated a fully humanized Dsg3 (hDSG3) murine model utilizing a hDsg3 transgenic animal crossed to the mDsg3 knockout line. Expression of hDsg3 in the mucosa rescues the mDsg3 knockout phenotype. Well-characterized mPV sera bind mucosal epithelia from the hDsg3 mice, but not mucosal tissues from WT mice, as detected by indirect immunofluorescence (IF). The majority of mPV sera preferentially recognize hDsg3 compared with mDsg3 by immunoprecipitation as well. Passive transfer of mPV IgG into adult hDsg3 mice, but not WT mice, induces suprabasilar acantholysis in mucosal tissues, thus confirming the pathogenicity of mPV anti-hDsg3 IgG in vivo. Human anti-hDsg3 antibodies are detected in perilesional mucosa as well as in sera of recipient mice by IF. These findings suggest that the Dsg3 epitopes targeted by pathogenic mPV IgG are human specific. This hDsg3 mouse model will be invaluable in studying the clinical transition from mPV to mcPV.

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Related in: MedlinePlus

Sera from mPV patients preferentially recognize hDsg3. Sera from well characterized mPV and mcPV patients were tested by indirect IF (a) using WT and hDsg3Tg mucosa with anti-human IgG4 FITC as the secondary antibody. Healthy control sera (Neg) and pemphigus foliaceus (PF) sera are included as negative and positive controls, respectively. Bar = 25um. (b) Immunoprecipitation was performed using either recombinant his tagged hDsg3 or mDsg3 and patient sera. Immunoprecipitated hDsg3 or mDsg3 was detected by anti-His HRP. All detected bands ran at the expected size of approximately 77kDa. Noncontiguous lanes run on the same gel are indicated by a white dashed line.
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Figure 4: Sera from mPV patients preferentially recognize hDsg3. Sera from well characterized mPV and mcPV patients were tested by indirect IF (a) using WT and hDsg3Tg mucosa with anti-human IgG4 FITC as the secondary antibody. Healthy control sera (Neg) and pemphigus foliaceus (PF) sera are included as negative and positive controls, respectively. Bar = 25um. (b) Immunoprecipitation was performed using either recombinant his tagged hDsg3 or mDsg3 and patient sera. Immunoprecipitated hDsg3 or mDsg3 was detected by anti-His HRP. All detected bands ran at the expected size of approximately 77kDa. Noncontiguous lanes run on the same gel are indicated by a white dashed line.

Mentions: Thus, hDsg3 is expressed in hDsg3Tg mice in a similar pattern to that of mDsg3 in WT mice. Furthermore, expression of endogenous Dsg1 and desmocollin 3 (Dsc3) is similar in WT and hDsg3Tg skin and mucosa, suggesting that introduction of hDsg3 does not alter Dsg1 or Dsc3 expression in the hDsg3Tg mice (Supplementary Figure 2 and Figure 4a).


Mucosal pemphigus vulgaris anti-Dsg3 IgG is pathogenic to the oral mucosa of humanized Dsg3 mice.

Culton DA, McCray SK, Park M, Roberts JC, Li N, Zedek DC, Anhalt GJ, Cowley DO, Liu Z, Diaz LA - J. Invest. Dermatol. (2015)

Sera from mPV patients preferentially recognize hDsg3. Sera from well characterized mPV and mcPV patients were tested by indirect IF (a) using WT and hDsg3Tg mucosa with anti-human IgG4 FITC as the secondary antibody. Healthy control sera (Neg) and pemphigus foliaceus (PF) sera are included as negative and positive controls, respectively. Bar = 25um. (b) Immunoprecipitation was performed using either recombinant his tagged hDsg3 or mDsg3 and patient sera. Immunoprecipitated hDsg3 or mDsg3 was detected by anti-His HRP. All detected bands ran at the expected size of approximately 77kDa. Noncontiguous lanes run on the same gel are indicated by a white dashed line.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4430403&req=5

Figure 4: Sera from mPV patients preferentially recognize hDsg3. Sera from well characterized mPV and mcPV patients were tested by indirect IF (a) using WT and hDsg3Tg mucosa with anti-human IgG4 FITC as the secondary antibody. Healthy control sera (Neg) and pemphigus foliaceus (PF) sera are included as negative and positive controls, respectively. Bar = 25um. (b) Immunoprecipitation was performed using either recombinant his tagged hDsg3 or mDsg3 and patient sera. Immunoprecipitated hDsg3 or mDsg3 was detected by anti-His HRP. All detected bands ran at the expected size of approximately 77kDa. Noncontiguous lanes run on the same gel are indicated by a white dashed line.
Mentions: Thus, hDsg3 is expressed in hDsg3Tg mice in a similar pattern to that of mDsg3 in WT mice. Furthermore, expression of endogenous Dsg1 and desmocollin 3 (Dsc3) is similar in WT and hDsg3Tg skin and mucosa, suggesting that introduction of hDsg3 does not alter Dsg1 or Dsc3 expression in the hDsg3Tg mice (Supplementary Figure 2 and Figure 4a).

Bottom Line: There are two major clinical subsets of pemphigus vulgaris (PV)-mucosal PV (mPV) and mucocutaneous PV (mcPV).The majority of mPV sera preferentially recognize hDsg3 compared with mDsg3 by immunoprecipitation as well.These findings suggest that the Dsg3 epitopes targeted by pathogenic mPV IgG are human specific.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

ABSTRACT
There are two major clinical subsets of pemphigus vulgaris (PV)-mucosal PV (mPV) and mucocutaneous PV (mcPV). The mPV subset exhibits anti-human desmoglein (Dsg) 3 autoantibodies that fail to recognize murine Dsg3 (mDsg3); thus, passive transfer experiments of mPV IgG into wild-type (WT) mice have been unsuccessful at inducing disease. We therefore generated a fully humanized Dsg3 (hDSG3) murine model utilizing a hDsg3 transgenic animal crossed to the mDsg3 knockout line. Expression of hDsg3 in the mucosa rescues the mDsg3 knockout phenotype. Well-characterized mPV sera bind mucosal epithelia from the hDsg3 mice, but not mucosal tissues from WT mice, as detected by indirect immunofluorescence (IF). The majority of mPV sera preferentially recognize hDsg3 compared with mDsg3 by immunoprecipitation as well. Passive transfer of mPV IgG into adult hDsg3 mice, but not WT mice, induces suprabasilar acantholysis in mucosal tissues, thus confirming the pathogenicity of mPV anti-hDsg3 IgG in vivo. Human anti-hDsg3 antibodies are detected in perilesional mucosa as well as in sera of recipient mice by IF. These findings suggest that the Dsg3 epitopes targeted by pathogenic mPV IgG are human specific. This hDsg3 mouse model will be invaluable in studying the clinical transition from mPV to mcPV.

Show MeSH
Related in: MedlinePlus