Analyzing pathogenic (double-stranded (ds) DNA-specific) plasma cells via immunofluorescence microscopy.
BACKGROUND
While protective plasma cells (PCs) are an important part of the individual's immune defense, autoreactive plasma cells such as dsDNA-specific plasma cells contribute to the pathogenesis of autoimmune diseases like systemic lupus erythematosus (SLE). However, the research on dsDNA-specific plasma cells was restricted to the ELISpot technique, with its limitations, as no other attempt for identification of dsDNA-reactive plasma cells had been successful.


METHODS
With improved fluorochrome labeling of dsDNA, removal of DNA aggregates, and enhanced blocking of unspecific binding, we were able to specifically detect dsDNA-reactive plasma cells by immunofluorescence microscopy.


RESULTS
Via this novel technique we were able to distinguish short-lived (SLPCs) and long-lived (LLPCs) autoreactive plasma cells, discriminate dsDNA-specific plasma cells according to their immunoglobulin class (IgG, IgM, and IgA) and investigate autoreactive (dsDNA) and vaccine-induced ovalbumin (Ova) plasma cells in parallel.


CONCLUSIONS
The detection of autoreactive dsDNA-specific plasma cells via immunofluorescence microscopy allows specific studies on pathogenic and protective plasma cell subsets and their niches, detailed evaluation of therapeutic treatments and therefore offers new possibilities for basic and clinical research.
Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany. oliver.winter@charite.de.|Department of Neonatology, Charité - Universitätsmedizin Berlin, Augustenburger Platz 1, 13353, Berlin, Germany. oliver.winter@charite.de.
Oliver
O
Winter
Department of Autoimmunology, Deutsches Rheuma-Forschungszentrum, Charitéplatz 1, 10117, Berlin, Germany. musiol89@gmx.net.
Stephanie
S
Musiol
Department of Autoimmunology, Deutsches Rheuma-Forschungszentrum, Charitéplatz 1, 10117, Berlin, Germany. fiorina@vollbio.de.
Melissa
M
Schablowsky
Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany. cheng@drfz.de.|Department of Autoimmunology, Deutsches Rheuma-Forschungszentrum, Charitéplatz 1, 10117, Berlin, Germany. cheng@drfz.de.
Qingyu
Q
Cheng
Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany. khodadadi@drfz.de.|Department of Autoimmunology, Deutsches Rheuma-Forschungszentrum, Charitéplatz 1, 10117, Berlin, Germany. khodadadi@drfz.de.
Laleh
L
Khodadadi
Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany. falk.hiepe@charite.de.
Falk
F
Hiepe
D000818:Animals; D000974:Antibodies, Antinuclear / Q000097:blood / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology; D004247:DNA / Q000276:immunology*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*; D010950:Plasma Cells / Q000276:immunology*
D000974:Antibodies, Antinuclear; D001324:Autoantigens; D004247:DNA