A comparative study of antibody expressions in primary biliary cirrhosis and autoimmune cholangitis using phage display

Hepatology. 2001 Sep;34(3):478-86. doi: 10.1053/jhep.2001.27013.

Abstract

Primary biliary cirrhosis (PBC) and autoimmune cholangitis (AIC) are serologic expressions of an autoimmune liver disease affecting biliary ductular cells. Previously we screened a phage-displayed random peptide library with polyclonal IgG from 2 Australian patients with PBC and derived peptides that identified a single conformational (discontinuous) epitope in the inner lipoyl domain of the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2), the characteristic autoantigen in PBC. Here we have used phage display to investigate the reactivity of PBC sera from 2 ethnically and geographically distinct populations, Japanese and Australian, and the 2 serologic expressions, PBC and AIC. Random 7-mer and 12-mer peptide libraries were biopanned with IgG from 3 Japanese patients with PBC and 3 with AIC who did not have anti-PDC-E2. The phage clones (phagotopes) obtained were tested by capture enzyme-linked immunosorbent assay (ELISA) for reactivity with affinity-purified anti-PDC-E2, and compared with those obtained from Australian patients with PBC. Peptide sequences of the derived phagotopes and sequences derived by biopanning with irrelevant antisera were aligned to develop a guide tree based on physicochemical similarity. Both Australian and Japanese PBC-derived phagotopes were distributed in branches of the guide tree that contained the peptide sequences MH and FV previously identified as part of an immunodominant conformational epitope of PDC-E2, indicating that epitope selection was not influenced by the racial origin of the PBC sera. Biopanning with either PBC or AIC-derived IgG yielded phagotopes that reacted with anti-PDC-E2 by capture ELISA, further establishing that there is a similar autoimmune targeting in PBC and AIC.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Aged
  • Algorithms
  • Antibodies / analysis*
  • Antigens, Nuclear*
  • Autoantigens / immunology
  • Autoimmune Diseases / immunology*
  • Cell Line
  • Cholangitis / immunology*
  • Dihydrolipoyllysine-Residue Acetyltransferase
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Fluorescent Antibody Technique
  • Humans
  • Liver Cirrhosis, Biliary / immunology*
  • Middle Aged
  • Nuclear Proteins / immunology
  • Peptide Library
  • Pyruvate Dehydrogenase Complex / immunology

Substances

  • Antibodies
  • Antigens, Nuclear
  • Autoantigens
  • Nuclear Proteins
  • Peptide Library
  • Pyruvate Dehydrogenase Complex
  • SP100 protein, human
  • Dihydrolipoyllysine-Residue Acetyltransferase