Anti-kelch-like 12 and Anti-Hexokinase 1: Novel Autoantibodies in Primary Biliary Cirrhosis

Liver Int. 2015 Feb;35(2):642-51. doi: 10.1111/liv.12690. Epub 2014 Oct 10.

Abstract

Background & aims: Using high-density human recombinant protein microarrays, we identified two potential biomarkers, kelch-like 12 (KLHL12) and hexokinase-1 (HK1), in primary biliary cirrhosis (PBC). The objective of this study was to determine the diagnostic value of anti-KLHL12/HK1 autoantibodies in PBC. Initial discovery used sera from 22 patients with PBC and 62 non-PBC controls. KLHL12 and HK1 proteins were then analysed for immunoglobulin reactivity by immunoblot and enzyme-linked immunosorbent assay (ELISA) in two independent cohorts of PBC and disease/healthy control patients.

Methods: Serum samples from 100 patients with PBC and 165 non-PBC disease controls were analysed by immunoblot and samples from 366 patients with PBC, 174 disease controls, and 80 healthy donors were tested by ELISA.

Results: Anti-KLHL12 and anti-HK1 antibodies were each detected more frequently in PBC compared with non-PBC disease controls (P < 0.001). Not only are both markers highly specific for PBC (≥95%) but they also yielded higher sensitivity than anti-gp210 and anti-sp100 antibodies. Combining anti-HK1 and anti-KLHL12 with available markers (MIT3, gp210 and sp100), increased the diagnostic sensitivity for PBC. Most importantly, anti-KLHL12 and anti-HK1 antibodies were present in 10-35% of anti-mitochondrial antibody (AMA)-negative PBC patients and adding these two biomarkers to conventional PBC assays dramatically improved the serological sensitivity in AMA-negative PBC from 55% to 75% in immunoblot and 48.3% to 68.5% in ELISA.

Conclusions: The addition of tests for highly specific anti-KLHL12 and anti-HK1 antibodies to AMA and ANA serological assays significantly improves efficacy in the clinical detection and diagnosis of PBC, especially for AMA-negative subjects.

Keywords: diagnostics; liver; serology.

Publication types

  • Clinical Trial
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantibodies* / blood
  • Autoantibodies* / immunology
  • Biomarkers / blood*
  • Enzyme-Linked Immunosorbent Assay
  • Hexokinase / immunology*
  • Humans
  • Immunoblotting
  • Liver Cirrhosis, Biliary / blood
  • Liver Cirrhosis, Biliary / immunology*
  • Microfilament Proteins / immunology*
  • Protein Array Analysis
  • Sensitivity and Specificity

Substances

  • Autoantibodies
  • Biomarkers
  • KLHL12 protein, human
  • Microfilament Proteins
  • anti-KLHL12 antibody, human
  • anti-hexokinase 1 antibody, human
  • HK1 protein, human
  • Hexokinase