TY - JOUR
T1 - Detection of IgG autoantibodies in the sera of patients with bullous and gestational pemphigoid
T2 - ELISA studies utilizing a baculovirus-encoded form of bullous pemphigoid antigen 2
AU - Haase, Claudia
AU - Büdinger, Lioba
AU - Borradori, Luca
AU - Yee, Carole
AU - Merk, Hans F.
AU - Yancey, Kim
AU - Hertl, Michael
N1 - Funding Information:
We thank Profs. Dr. H. Pullmann and E. Haneke and Drs. A. Schattenkirchner, A. Mira, and T. Zollner for providing BP sera for this study. This study was supported by grant He 1604/5—1 from the Deutsche Forschungsgemeinschaft, Bonn, Germany. L.B. has been supported by a grant from the Swiss National Foundation for Scientific Research.
PY - 1998
Y1 - 1998
N2 - Autoantibodies against the extracellular domain of bullous pemphigoid antigen 2 (BPAG2) are thought to play a key role in the pathogenesis of bullous pemphigoid and their detection may thus be of diagnostic and prognostic value. The aim of this study was to develop a standardized enzyme- linked immunosorbent assay utilizing the baculovirus-derived protein BV13 (extracellular domain of BPAG2 devoid of 68 amino acids at the C terminus linked to glutathione-S-transferase and 6x His tag) to detect BPAG2-specific autoantibodies. For the enzyme-linked immunosorbent assay, nickel agarose affinity-purified BV13 protein was incubated with sera from patients with bullous pemphigoid (n = 39), gestational pemphigoid (n = 10), and pemphigus vulgaris/pemphigus foliaceus (PV/PF; n = 15), or normal human sera (NHS; n = 18). Nickel affinity-purified proteins from wild-type baculovirus-infected insect cells served as a control. A positive enzyme-linked immunosorbent assay value was defined as reactivity (OD(BV13) -OD(WT)) > mean reactivity + 1 SD of the negative control sera (PV/PF; NHS). Thirty-five of 39 bullous pemphigoid sera and 10 of 10 gestational pemphigoid sera were reactive to BPAG2 compared with none of 15 PV/PF sera and one of 18 NHS (sensitivity, 91.8%; specificity, 97%). Of 16 BPAG2-reactive sera in the enzyme-linked immunosorbent assay, only six were BPAG2-reactive in the western blot, whereas 14 sera immunoprecipitated BPAG2 from extracts of epidermal keratinocytes. The enzyme-linked immunosorbent assay utilizing an eukaryotic BPAG2 protein thus seems to be highly sensitive and specific in the detection of BPAG2-specific antibodies and, hence, may be useful in the diagnosis of bullous autoimmune diseases, such, as bullous pemphigoid and gestational pemphigoid.
AB - Autoantibodies against the extracellular domain of bullous pemphigoid antigen 2 (BPAG2) are thought to play a key role in the pathogenesis of bullous pemphigoid and their detection may thus be of diagnostic and prognostic value. The aim of this study was to develop a standardized enzyme- linked immunosorbent assay utilizing the baculovirus-derived protein BV13 (extracellular domain of BPAG2 devoid of 68 amino acids at the C terminus linked to glutathione-S-transferase and 6x His tag) to detect BPAG2-specific autoantibodies. For the enzyme-linked immunosorbent assay, nickel agarose affinity-purified BV13 protein was incubated with sera from patients with bullous pemphigoid (n = 39), gestational pemphigoid (n = 10), and pemphigus vulgaris/pemphigus foliaceus (PV/PF; n = 15), or normal human sera (NHS; n = 18). Nickel affinity-purified proteins from wild-type baculovirus-infected insect cells served as a control. A positive enzyme-linked immunosorbent assay value was defined as reactivity (OD(BV13) -OD(WT)) > mean reactivity + 1 SD of the negative control sera (PV/PF; NHS). Thirty-five of 39 bullous pemphigoid sera and 10 of 10 gestational pemphigoid sera were reactive to BPAG2 compared with none of 15 PV/PF sera and one of 18 NHS (sensitivity, 91.8%; specificity, 97%). Of 16 BPAG2-reactive sera in the enzyme-linked immunosorbent assay, only six were BPAG2-reactive in the western blot, whereas 14 sera immunoprecipitated BPAG2 from extracts of epidermal keratinocytes. The enzyme-linked immunosorbent assay utilizing an eukaryotic BPAG2 protein thus seems to be highly sensitive and specific in the detection of BPAG2-specific antibodies and, hence, may be useful in the diagnosis of bullous autoimmune diseases, such, as bullous pemphigoid and gestational pemphigoid.
KW - Autoimmunity
KW - B lymphocytes
KW - Hemidesmosome
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UR - http://www.scopus.com/inward/citedby.url?scp=0031917777&partnerID=8YFLogxK
U2 - 10.1046/j.1523-1747.1998.00126.x
DO - 10.1046/j.1523-1747.1998.00126.x
M3 - Article
C2 - 9506450
AN - SCOPUS:0031917777
SN - 0022-202X
VL - 110
SP - 282
EP - 286
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 3
ER -