Identification of target antigen for SLA/LP autoantibodies in autoimmune hepatitis.
BACKGROUND
Autoantibodies are a hallmark of autoimmune hepatitis, but most are not disease specific. Autoantibodies to soluble liver antigen (SLA) and to liver and pancreas antigen (LP) have been described as disease specific, occurring in about 30% of all patients with autoimmune hepatitis, but no standardised assays are available. Methods We tested 2000 serum samples from patients with various liver diseases and controls for SLA autoantibodies by inhibition ELISA. Serum samples positive for SLA antibodies were used for immunoscreening of cDNA expression libraries. Identified clones were tested against a panel of serum samples positive for SLA and LP autoantibodies and control serum samples, and the epitope mapped by deletion mutants and exonuclease digestion.


RESULTS
SLA and LP autoantibodies were identical. Of 2000 serum samples screened, 35 were positive for SLA autoantibodies. These positive samples came from patients with autoimmune hepatitis; three from patients with an overlap syndrome (primary biliary cirrhosis and secondary autoimmune hepatitis). Expression cloning and absorption experiments identified a 422 aminoacid protein present in two splice variants as the sole target antigen. Aminoacids 371-409 were critical for immune recognition.


CONCLUSIONS
The identified cDNA encodes the primary target antigen of SLA/LP autoantibodies. The SLA/LP antigen has a previously unknown aminoacid sequence, and presumably codes for an unindentified enzyme, suggested to be UGA-suppressor tRNA-associated protein. SLA/LP autoantibodies are disease specific and recognise a dominant epitope, suggesting a specific antigen-driven immune response. Identification of the SLA/LP target antigen will allow establishment of a reliable, widely available diagnostic assay. Furthermore, its role in the pathogenesis of autoimmune hepatitis can now be studied.
Wies
I
I
Department of Medicine, Johannes Gutenberg University, Mainz, Germany.
Brunner
S
S
Henninger
J
J
Herkel
J
J
Kanzler
S
S
Meyer zum Büschenfelde
K H
KH
Lohse
A W
AW
D001323:Autoantibodies / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D018076:DNA, Complementary / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D019693:Hepatitis, Autoimmune / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D006801:Humans; D008099:Liver / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology; D008969:Molecular Sequence Data; D010179:Pancreas / Q000276:immunology*; D017385:Sequence Homology
D001323:Autoantibodies; D001324:Autoantigens; D018076:DNA, Complementary; D000939:Epitopes; C046431:liver antigen LA-1