Characterizing the proteome of bullous pemphigoid blister fluid utilizing tandem mass tag labeling coupled with LC–MS/MS

Farzan Solimani, Dario Didona, Jing Li, Lei Bao, Payal M. Patel, Giulia Gasparini, Khalaf Kridin, Emanuele Cozzani, Michael Hertl, Kyle T. Amber

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Bullous pemphigoid is an autoimmune blistering disease caused by autoantibodies against components of the cutaneous basement membrane zone. Autoantibodies lead to complement-dependent and -independent inflammation and blistering. Blister fluid is a valuable biologic resource, as it provides insight into both systemic and local microenvironment responses. Here, we utilized liquid chromatography with tandem mass spectrometry to characterize the bullous pemphigoid blister fluid proteome. We then depleted exosomes to better understand the exosomal versus non-exosomal proteome. We identified 339 proteins in the blister fluid of bullous pemphigoid patients. Gene ontology demonstrated enrichment of several key biologic processes including innate immune response, neutrophil degranulation, platelet degranulation, and complement activation. Exosome depletion resulted in a significant decrease in normalized reporter intensities of 192 proteins, consistent with our observation of a large number of exosomal proteins found in the blister fluid. We then compared the bullous pemphigoid blister fluid proteome to prior proteomic datasets in suction blister fluid, snake bites, and thermal burns, identifying 76 proteins unique to bullous pemphigoid. These include major basic protein, eosinophil peroxidase, galectin-10, and the immunoglobulin epsilon heavy constant region, consistent with tissue eosinophilia. We lastly validated several previously reported blister fluid exosomal components. Blister fluid in bullous pemphigoid contains a mixture of numerous biologic processes. While many of these processes are shared with blistering from alternative causes, we have identified several notable features unique to bullous pemphigoid.

Original languageEnglish
Pages (from-to)921-928
Number of pages8
JournalArchives of Dermatological Research
Volume314
Issue number9
DOIs
StatePublished - Nov 2022
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

Funding

Lei Bao received financial support from the Albert H. and Mary Jane Slepyan Endowed Fellowship. Farzan Soliamni and Michael Hertl were funded by grants from the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG); FOR 2497. The authors thank Creative Proteomics (Shirley, NY) for assistance with LC–MS/MS. Lei Bao received financial support from the Albert H. and Mary Jane Slepyan Endowed Fellowship. Farzan Soliamni and Michael Hertl were funded by grants from the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG); FOR 2497.

FundersFunder number
Creative Proteomics
California Department of Fish and Game
Deutsche ForschungsgemeinschaftFOR 2497

    Keywords

    • Autoimmune blistering disorder
    • Blister fluid
    • Pemphigoid
    • Proteomics
    • Quantitative proteomics

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