Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 282-286 |
| Number of pages | 5 |
| Journal | Journal of Investigative Dermatology |
| Volume | 110 |
| Issue number | 3 |
| DOIs | |
| State | Published - 1998 |
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Keywords
- Autoimmunity
- B lymphocytes
- Hemidesmosome
ASJC Scopus subject areas
- Dermatology
Cite this
Detection of IgG autoantibodies in the sera of patients with bullous and gestational pemphigoid : ELISA studies utilizing a baculovirus-encoded form of bullous pemphigoid antigen 2. / Haase, Claudia; Büdinger, Lioba; Borradori, Luca; Yee, Carole; Merk, Hans F.; Yancey, Kim; Hertl, Michael.
In: Journal of Investigative Dermatology, Vol. 110, No. 3, 1998, p. 282-286.Research output: Contribution to journal › Article
}
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
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
UR - http://www.scopus.com/inward/record.url?scp=0031917777&partnerID=8YFLogxK
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
VL - 110
SP - 282
EP - 286
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
SN - 0022-202X
IS - 3
ER -