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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.
J Invest Dermatol. 2003 Mar; 120(3):372-8.JI

Abstract

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.

Authors+Show Affiliations

Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12603849

Citation

Leyendeckers, Heike, et al. "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." The Journal of Investigative Dermatology, vol. 120, no. 3, 2003, pp. 372-8.
Leyendeckers H, Tasanen K, Bruckner-Tuderman L, et al. 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. J Invest Dermatol. 2003;120(3):372-8.
Leyendeckers, H., Tasanen, K., Bruckner-Tuderman, L., Zillikens, D., Sitaru, C., Schmitz, J., & Hunzelmann, N. (2003). 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. The Journal of Investigative Dermatology, 120(3), 372-8.
Leyendeckers H, et al. 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. J Invest Dermatol. 2003;120(3):372-8. PubMed PMID: 12603849.
* Article titles in AMA citation format should be in sentence-case
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, PY - 2003/2/27/pubmed PY - 2003/5/13/medline PY - 2003/2/27/entrez SP - 372 EP - 8 JF - The Journal of investigative dermatology JO - J Invest Dermatol VL - 120 IS - 3 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. SN - 0022-202X UR - https://www.unboundmedicine.com/medline/citation/12603849/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_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-202X(15)30185-8 DB - PRIME DP - Unbound Medicine ER -