Discrimination of agonist and antagonist forms of CXCL10 in biological samples

Clin Exp Immunol. 2012 Jan;167(1):137-48. doi: 10.1111/j.1365-2249.2011.04488.x.

Abstract

The ready access to commercially available multiplex assays and the importance of inflammation in disease pathogenesis has resulted in an abundance of studies aimed at identifying surrogate biomarkers for different clinically important questions. Establishing a link between a biomarker and disease pathogenesis, however, is quite complex, and in some instances this complexity is compounded by post-translational modifications and the use of immunoassays that do not always discriminate between the different forms of the same protein. Herein, we provide a detailed description of an assay system that has been established to discriminate the agonist form of CXCL10 from the NH(2) -terminal truncated form of the molecule generated by dipeptidylpeptidase IV (DPP4) cleavage. We demonstrate the utility of this assay system for monitoring agonist and antagonist forms of CXCL10 in culture supernatant, patient plasma and urine samples. Given the important role of CXCL10 in chronic inflammatory diseases and its suggested role as a predictive marker in managing patients with chronic hepatitis C, asthma, atopic dermatitis, transplantation, tuberculosis, kidney injury, cancer and other diseases, we believe that our method will be of general interest to the research and medical community.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies, Monoclonal / immunology
  • Biomarkers
  • Body Fluids / chemistry
  • Carcinoma, Transitional Cell / urine
  • Chemokine CXCL10 / analysis*
  • Chemokine CXCL10 / immunology
  • Culture Media, Conditioned / chemistry
  • Dipeptidyl Peptidase 4 / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Hepatitis C, Chronic / blood
  • Humans
  • Immunoenzyme Techniques / methods*
  • Inflammation
  • Male
  • Middle Aged
  • Neoplasm Proteins / urine
  • Peptide Fragments / analysis
  • Peptide Fragments / immunology
  • Protein Isoforms / analysis
  • Protein Isoforms / immunology
  • Protein Processing, Post-Translational
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / analysis
  • Urinary Bladder Neoplasms / urine

Substances

  • Antibodies, Monoclonal
  • Biomarkers
  • CXCL10 protein, human
  • Chemokine CXCL10
  • Culture Media, Conditioned
  • Neoplasm Proteins
  • Peptide Fragments
  • Protein Isoforms
  • Recombinant Fusion Proteins
  • DPP4 protein, human
  • Dipeptidyl Peptidase 4