TY - JOUR
T1 - Memory B cells specific for the NC16A domain of the 180 kDa bullous pemphigoid autoantigen can be detected in peripheral blood of bullous pemphigoid patients and induced in vitro to synthesize autoantibodies
AU - Leyendeckers, Heike
AU - Tasanen, Kaisa
AU - Bruckner-Tuderman, Leena
AU - Zillikens, Detlef
AU - Sitaru, Cassian
AU - Schmitz, Jürgen
AU - Hunzelmann, Nicolas
N1 - Funding Information:
We are grateful to Drs R. Zubler and E. Butcher for generous gifts of cell lines and antibodies. This study was supported by the Bundesministerium für Bildung, Wissenschaft, Forschung und Technologie, the Deutsche Forschungsgemeinschaft (DFG Hu 446/3-1), the Köln Fortune program and also by grants from the Academy of Finland, the Alexander von Humboldt Foundation and the Oulu University Hospital.
PY - 2003/3/1
Y1 - 2003/3/1
N2 - Bullous pemphigoid is a subepidermal blistering disease characterized by the synthesis of autoantibodies against the 180 kDa and the 230 kDa bullous pemphigoid antigens. Whether autoimmunity is also reflected by the presence of circulating autoantigen-specific memory B cells is still a matter of debate. We used a new assay combining two-step immunomagnetic enrichment with multiparameter flow cytometry to detect and characterize bullous pemphigoid 180 kDa-specific IgG+ B cells in blood of bullous pemphigoid patients. In a first magnetic separation, B cells were isolated from peripheral blood mononuclear cells using releasable microbeads conjugated to a CD19 antibody. From pre-enriched B cells, bullous pemphigoid 180 kDa-specific cells were then positively selected using microbeads directly conjugated with a recombinant N-terminal fragment of the bullous pemphigoid 180 kDa ectodomain, containing the noncollagenous 16A domain, which was recently shown to harbor major epitopes of autoantibodies in bullous pemphigoid sera. Noncollagenous 16A domain-specific IgG+ B cells were detectable in blood of most, if not all patients with serum autoantibodies against the noncollagenous 16A domain. The specificity of the cells was confirmed by in vitro differentiation into antibody-forming cells and analysis of the culture supernatant for the presence of noncollagenous 16A domain-specific IgG antibodies. All noncollagenous 16A domain-specific IgG+ B cells showed a clear memory immunophenotype. Noncollagenous 16A domain-specific IgG+ memory B cells may be crucial for continuous noncollagenous 16A domain-specific autoantibody production and/or play a part as antigen-presenting cells for priming and restimulation of bullous pemphigoid 180 kDa-specific T helper cells.
AB - Bullous pemphigoid is a subepidermal blistering disease characterized by the synthesis of autoantibodies against the 180 kDa and the 230 kDa bullous pemphigoid antigens. Whether autoimmunity is also reflected by the presence of circulating autoantigen-specific memory B cells is still a matter of debate. We used a new assay combining two-step immunomagnetic enrichment with multiparameter flow cytometry to detect and characterize bullous pemphigoid 180 kDa-specific IgG+ B cells in blood of bullous pemphigoid patients. In a first magnetic separation, B cells were isolated from peripheral blood mononuclear cells using releasable microbeads conjugated to a CD19 antibody. From pre-enriched B cells, bullous pemphigoid 180 kDa-specific cells were then positively selected using microbeads directly conjugated with a recombinant N-terminal fragment of the bullous pemphigoid 180 kDa ectodomain, containing the noncollagenous 16A domain, which was recently shown to harbor major epitopes of autoantibodies in bullous pemphigoid sera. Noncollagenous 16A domain-specific IgG+ B cells were detectable in blood of most, if not all patients with serum autoantibodies against the noncollagenous 16A domain. The specificity of the cells was confirmed by in vitro differentiation into antibody-forming cells and analysis of the culture supernatant for the presence of noncollagenous 16A domain-specific IgG antibodies. All noncollagenous 16A domain-specific IgG+ B cells showed a clear memory immunophenotype. Noncollagenous 16A domain-specific IgG+ memory B cells may be crucial for continuous noncollagenous 16A domain-specific autoantibody production and/or play a part as antigen-presenting cells for priming and restimulation of bullous pemphigoid 180 kDa-specific T helper cells.
UR - http://www.scopus.com/inward/record.url?scp=0344838391&partnerID=8YFLogxK
U2 - 10.1046/j.1523-1747.2003.12071.x
DO - 10.1046/j.1523-1747.2003.12071.x
M3 - Journal articles
C2 - 12603849
AN - SCOPUS:0344838391
SN - 0022-202X
VL - 120
SP - 372
EP - 378
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 3
ER -