Abstract
Reactivity of sera from patients with primary biliary cirrhosis (PBC) with a 60 kDa component of nuclear pore complexes (NPCs), purified by affinity chromatography on wheat-germ agglutinin (WGA)-Sepharose, was previously detected. Recently, clinical significance of the anti-NPC antibodies in PBC became evident. In the light of recent reports, indicating the correlation of the anti-NPC antibodies with severity and progression of the disease, the characterization of the reactive antigens is becoming essential in the clinical management of patients with PBC. Since accurate autoantibody detection represents one of the fundamental requirements for a reliable testing, we have generated a human recombinant p62 protein and validated an immunoprecipitation assay for the detection of anti-p62. We also demonstrated that the generated human recombinant p62 nucleoporin was modified by N-acetylglucosamine residues. More than50%of tested PBC sera precipitated 35S-radioactively labeled p62 recombinant nucleoporin and 40% recognized this recombinant antigen by immunoblotting. We compared the reactivity of PBC sera with rat and human nucleoporin. The incidence of anti-p62 nucleoporin positive PBC sera increased by 15% when human recombinant antigen was used. The titer of autoantibodies in p62-positive PBC samples strongly varied. Preadsorption of the PBC sera with p62 recombinant protein completely abolished their reactivity with the antigen. In conclusion, this study unequivocally proves that autoantibodies reacting with the 60 kDa component of NPCs target p62 nucleoporin and, more importantly, provide a better antigen source for future evaluations of the clinical role of anti-p62 in PBC.
| Original language | English |
|---|---|
| Pages (from-to) | 27-37 |
| Number of pages | 11 |
| Journal | Journal of Cellular Biochemistry |
| Volume | 104 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - May 1 2008 |
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Keywords
- ANA
- Autoimmunity
- gp210 glycoprotein
- Human recombinant p62 nucleoporin
- Nuclear envelope
- Nuclear pore complexes
- p62 nucleoporin
ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Molecular Biology
Cite this
Characterization of the antibodies to p62 nucleoporin in primary biliary cirrhosis using human recombinant antigen. / Wȩsierska-Ga̧dek, Józefa; Klima, Anna; Ranftler, Carmen; Komina, Oxana; Hanover, John; Invernizzi, Pietro; Penner, Edward.
In: Journal of Cellular Biochemistry, Vol. 104, No. 1, 01.05.2008, p. 27-37.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of the antibodies to p62 nucleoporin in primary biliary cirrhosis using human recombinant antigen
AU - Wȩsierska-Ga̧dek, Józefa
AU - Klima, Anna
AU - Ranftler, Carmen
AU - Komina, Oxana
AU - Hanover, John
AU - Invernizzi, Pietro
AU - Penner, Edward
PY - 2008/5/1
Y1 - 2008/5/1
N2 - Reactivity of sera from patients with primary biliary cirrhosis (PBC) with a 60 kDa component of nuclear pore complexes (NPCs), purified by affinity chromatography on wheat-germ agglutinin (WGA)-Sepharose, was previously detected. Recently, clinical significance of the anti-NPC antibodies in PBC became evident. In the light of recent reports, indicating the correlation of the anti-NPC antibodies with severity and progression of the disease, the characterization of the reactive antigens is becoming essential in the clinical management of patients with PBC. Since accurate autoantibody detection represents one of the fundamental requirements for a reliable testing, we have generated a human recombinant p62 protein and validated an immunoprecipitation assay for the detection of anti-p62. We also demonstrated that the generated human recombinant p62 nucleoporin was modified by N-acetylglucosamine residues. More than50%of tested PBC sera precipitated 35S-radioactively labeled p62 recombinant nucleoporin and 40% recognized this recombinant antigen by immunoblotting. We compared the reactivity of PBC sera with rat and human nucleoporin. The incidence of anti-p62 nucleoporin positive PBC sera increased by 15% when human recombinant antigen was used. The titer of autoantibodies in p62-positive PBC samples strongly varied. Preadsorption of the PBC sera with p62 recombinant protein completely abolished their reactivity with the antigen. In conclusion, this study unequivocally proves that autoantibodies reacting with the 60 kDa component of NPCs target p62 nucleoporin and, more importantly, provide a better antigen source for future evaluations of the clinical role of anti-p62 in PBC.
AB - Reactivity of sera from patients with primary biliary cirrhosis (PBC) with a 60 kDa component of nuclear pore complexes (NPCs), purified by affinity chromatography on wheat-germ agglutinin (WGA)-Sepharose, was previously detected. Recently, clinical significance of the anti-NPC antibodies in PBC became evident. In the light of recent reports, indicating the correlation of the anti-NPC antibodies with severity and progression of the disease, the characterization of the reactive antigens is becoming essential in the clinical management of patients with PBC. Since accurate autoantibody detection represents one of the fundamental requirements for a reliable testing, we have generated a human recombinant p62 protein and validated an immunoprecipitation assay for the detection of anti-p62. We also demonstrated that the generated human recombinant p62 nucleoporin was modified by N-acetylglucosamine residues. More than50%of tested PBC sera precipitated 35S-radioactively labeled p62 recombinant nucleoporin and 40% recognized this recombinant antigen by immunoblotting. We compared the reactivity of PBC sera with rat and human nucleoporin. The incidence of anti-p62 nucleoporin positive PBC sera increased by 15% when human recombinant antigen was used. The titer of autoantibodies in p62-positive PBC samples strongly varied. Preadsorption of the PBC sera with p62 recombinant protein completely abolished their reactivity with the antigen. In conclusion, this study unequivocally proves that autoantibodies reacting with the 60 kDa component of NPCs target p62 nucleoporin and, more importantly, provide a better antigen source for future evaluations of the clinical role of anti-p62 in PBC.
KW - ANA
KW - Autoimmunity
KW - gp210 glycoprotein
KW - Human recombinant p62 nucleoporin
KW - Nuclear envelope
KW - Nuclear pore complexes
KW - p62 nucleoporin
UR - http://www.scopus.com/inward/record.url?scp=46449135226&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=46449135226&partnerID=8YFLogxK
U2 - 10.1002/jcb.21595
DO - 10.1002/jcb.21595
M3 - Article
C2 - 17960595
AN - SCOPUS:46449135226
VL - 104
SP - 27
EP - 37
JO - Journal of Cellular Biochemistry
JF - Journal of Cellular Biochemistry
SN - 0730-2312
IS - 1
ER -