Antibodies against keratinocyte antigens other than desmogleins 1 and 3 can induce pemphigus vulgaris–like lesions
Vu Thuong Nguyen, … , Mark R. Pittelkow, Sergei A. Grando
Vu Thuong Nguyen, … , Mark R. Pittelkow, Sergei A. Grando
Published December 15, 2000
Citation Information: J Clin Invest. 2000;106(12):1467-1479. https://doi.org/10.1172/JCI10305.
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Article

Antibodies against keratinocyte antigens other than desmogleins 1 and 3 can induce pemphigus vulgaris–like lesions

Abstract

Pemphigus is an autoimmune disease of skin adhesion associated with autoantibodies against a number of keratinocyte antigens, such as the adhesion molecules desmoglein (Dsg) 1 and 3 and acetylcholine receptors. The notion that anti-Dsg antibodies alone are responsible for blisters in patients with pemphigus vulgaris (PV) stems from the ability of rDsg1 and rDsg3 to absorb antibodies that cause PV-like skin blisters in neonatal mice. Here, we demonstrate that PV IgGs eluted from rDsg1-Ig-His and rDsg3-Ig-His show similar antigenic profiles, including the 38-, 43-, 115-, and 190-kDa keratinocyte proteins and a non–Dsg 3 130-kDa polypeptide present in keratinocytes from Dsg 3 knockout mouse. We injected into Dsg 3–lacking mice the PV IgGs that did not cross-react with the 160-kDa Dsg 1 or its 45-kDa immunoreactive fragment and that showed no reactivity with recombinant Dsg 1. We used both the Dsg3null mice with a targeted mutation of the Dsg3 gene and the “balding” Dsg3bal/Dsg3bal mice that carry a spontaneous null mutation in Dsg3. These PV IgGs caused gross skin blisters with PV-like suprabasal acantholysis and stained perilesional epidermis in a fishnet-like pattern, indicating that the PV phenotype can be induced without anti–Dsg 3 antibody. The anti–Dsg 1 antibody also was not required, as its presence in PV IgG does not alter the PV-like phenotype in skin organ cultures and because pemphigus foliaceus IgGs produce a distinct phenotype in Dsg3null mice. Therefore, mucocutaneous lesions in PV patients could be caused by non-Dsg antibodies.

Authors

Vu Thuong Nguyen, Assane Ndoye, Leonard D. Shultz, Mark R. Pittelkow, Sergei A. Grando

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Figure 6

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The profiles of the PV IgGs absorbed by recombinant Dsg 1 and Dsg 3 bacu...
The profiles of the PV IgGs absorbed by recombinant Dsg 1 and Dsg 3 baculoproteins. Western blots of protein extracts of normal human keratinocytes resolved by 7.5% SDS-PAGE and stained with PV3 IgG affinity-purified on rDsg3-His (lane 1), and that of Dsg3null keratinocytes stained with PV3 IgG affinity purified on rDsg3-Ig-His (lane 5), rDsg1-Ig-His (lane 4) or on the Fc IgG column (lane 9). Binding of PV3 IgGs to the immunoblotting membranes was visualized using secondary, HRP-conjugated goat anti-human IgG antibodies. Lane 6 shows a single protein band with apparent Mr of 190 kDa among SDS-PAGE–resolved Dsg 3–positive keratinocyte proteins from BALB/c mouse. This band was visualized by the PV3 IgG that was eluted from the 190-kDa area of the Western blot of Dsg3null keratinocyte proteins stained with the PV3 IgG adsorbed on rDsg3-Ig-His. Note: Only a 190-kDa but not 130- or 160-kDa band or any other keratinocyte protein was visualized, indicating that the antibody targeting the 190-kDa protein is a unique one, as it does not recognize the 130-kDa Dsg 3 or 160-kDa Dsg 1. Lanes 2, 3, and 7 are the negative controls omitting primary antibody. The Dsg3null keratinocyte protein extract in lane 8 was blotted with normal human IgG-affinity purified on rDsg3-Ig-His. The lack of multiple bands in lane 8 as well as complete absence of specific staining in lanes 3 and 7 indicate that there were no nonspecific cross reactivities of human or goat IgGs with murine epidermal proteins. The positions of relative molecular mass markers run in parallel lanes of each blot are shown to the left of the respective blot. The apparent relative molecular mass of keratinocyte protein bands visualized due to PV antibody binding is shown to the right of lanes 2 and 5 in the columns designated Mr.