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BMC Immunol. 2012; 13: 16.
Published online Apr 4, 2012. doi:  10.1186/1471-2172-13-16
PMCID: PMC3368718
Prevalence of collagen VII-specific autoantibodies in patients with autoimmune and inflammatory diseases
Emilia Licarete,1,2 Susanne Ganz,1 Martin J Recknagel,3 Giovanni Di Zenzo,4 Takashi Hashimoto,5 Michael Hertl,6 Giovanna Zambruno,4 Gheorghe Hundorfean,7 Jonas Mudter,7 Markus F Neurath,7 Leena Bruckner-Tuderman,1,8 and Cassian Sitarucorresponding author1,8
1Department of Dermatology, University of Freiburg, Hauptstr. 7, Freiburg 79104, Germany
2Department of Experimental Biology and Molecular Biology Center, Institute for Interdisciplinary Research on Bio-Nano-Sciences, Babes-Bolyai University Cluj-Napoca, Cluj-Napoca, Romania
3Faculty of Biology, Genetics and Experimental Bioinformatics Group, University of Freiburg, Freiburg, Germany
4Molecular and Cell Biology Laboratory, IDI-IRCCS, Rome, Italy
5Department of Dermatology, Kurume University, 67 Asahimachi, Kurume, Fukuoka 830-0011, Japan
6Department of Dermatology and Allergology, University of Marburg, Baldingerdstraße, Marburg 33043, Germany
7University of Erlangen-Nuremberg, Medical Clinic I, Erlangen, Ulmenweg 18, Erlangen 91054, Germany
8Centre for Biological Signalling Studies (bioss), University of Freiburg, Freiburg, Germany
corresponding authorCorresponding author.
Emilia Licarete: emilia_licarete/at/yahoo.com; Susanne Ganz: ganz.susanne/at/googlemail.com; Martin J Recknagel: martin.recknagel/at/web.de; Giovanni Di Zenzo: g.dizenzo/at/idi.it; Takashi Hashimoto: hashimot/at/med.kurume-u.ac.jp; Michael Hertl: hertl/at/med.uni-marburg.de; Giovanna Zambruno: g.zambruno/at/idi.it; Gheorghe Hundorfean: Gheorghe.Hundorfean/at/uk-erlangen.de; Jonas Mudter: Jonas.Mudter/at/uk-erlangen.de; Markus F Neurath: markus.neurath/at/uk-erlangen.de; Leena Bruckner-Tuderman: leena.bruckner-tuderman/at/uniklinik-freiburg.de; Cassian Sitaru: cassian/at/mail.sitaru.eu
Received November 25, 2011; Accepted April 4, 2012.
Abstract
Background
Autoimmunity to collagen VII is typically associated with the skin blistering disease epidermolysis bullosa acquisita (EBA), but also occurs occasionally in patients with systemic lupus erythematosus or inflammatory bowel disease. The aim of our present study was to develop an accurate immunoassay for assessing the presence of autoantibodies against collagen VII in large cohorts of patients and healthy donors.
Methods
Based on in silico antigenic analysis and previous wetlab epitope mapping data, we designed a chimeric collagen VII construct containing all collagen VII epitopes with higher antigenicity. ELISA was performed with sera from patients with EBA (n = 50), Crohn's disease (CD, n = 50), ulcerative colitis (UC, n = 50), bullous pemphigoid (BP, n = 76), and pemphigus vulgaris (PV, n = 42) and healthy donors (n = 245).
Results
By ELISA, the receiver operating characteristics analysis yielded an area under the curve of 0.98 (95% CI: 0.9638-1.005), allowing to set the cut-off at 0.32 OD at a calculated specificity of 98% and a sensitivity of 94%. Running the optimized test showed that serum IgG autoantibodies from 47 EBA (94%; 95% CI: 87.41%-100%), 2 CD (4%; 95% CI: 0%-9.43%), 8 UC (16%; 95% CI: 5.8%-26%), 2 BP (2.63%; 95% CI: 0%-6.23%), and 4 PV (9.52%; 95% CI: 0%-18.4%) patients as well as from 4 (1.63%; 95% CI: 0%-3.21%) healthy donors reacted with the chimeric protein. Further analysis revealed that in 34%, 37%, 16% and 100% of sera autoantibodies of IgG1, IgG2, IgG3, and IgG4 isotype, respectively, recognized the recombinant autoantigen.
Conclusions
Using a chimeric protein, we developed a new sensitive and specific ELISA to detect collagen specific antibodies. Our results show a low prevalence of collagen VII-specific autoantibodies in inflammatory bowel disease, pemphigus and bullous pemphigoid. Furthermore, we show that the autoimmune response against collagen VII is dominated by IgG4 autoantibodies. The new immunoassay should prove a useful tool for clinical and translational research and should improve the routine diagnosis and disease monitoring in diseases associated with collagen VII-specific autoimmunity.
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