Quantitative assays for anti-aquaporin-4 antibody with subclass analysis in neuromyelitis optica

Mult Scler. 2012 Nov;18(11):1541-51. doi: 10.1177/1352458512443917. Epub 2012 Apr 23.

Abstract

Background: To clarify the clinical relevance of anti-aquaporin-4 (anti-AQP4) antibody titers and immunoglobulin (IgG) subclass.

Methods: Using a bridging enzyme-linked immunosorbent assay (ELISA), a flow cytometric assay (FCMA) and an immunofluorescence assay (IFA) for anti-AQP4 antibodies, sera from 142 patients with multiple sclerosis (MS) as defined by the McDonald criteria (2005), 29 with neuromyelitis optica (NMO) who fulfilled the 1999 criteria, 19 with recurrent and/or longitudinally extensive myelitis (RM/LM), 86 with other non-inflammatory neurological diseases (OND) and 28 healthy controls (HC) were studied.

Results: Anti-AQP4 antibody positivity rates by IFA, FCMA, and ELISA were 41.4%, 51.7% and 48.3%, respectively, in NMO (1999) patients, and 0% in the OND and HC groups. Twenty-six MS patients (18.3%) were positive for the antibody; 17 met the 2006 NMO criteria, including positivity for anti-AQP4 antibody, and five had longitudinally extensive myelitis (LM). Among the cases with anti-AQP4 antibody detected by FCMA, IgG1, 2, 3, and 4 anti-AQP4 antibodies were found in 97.8%, 37.0%, 6.5% and 6.5% respectively. There was no association of either antibody positivity or level of anti-AQP4 antibody IgG subclasses with clinical parameters after adjustment of p values for multiple comparisons.

Conclusions: FCMA and bridging ELISA are useful for detecting and quantifying anti-AQP4 antibodies.

Publication types

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

MeSH terms

  • Aquaporin 4 / genetics
  • Aquaporin 4 / immunology*
  • Autoantibodies / blood*
  • Autoantibodies / classification
  • Biomarkers / blood
  • Case-Control Studies
  • Chi-Square Distribution
  • Enzyme-Linked Immunosorbent Assay*
  • Flow Cytometry*
  • Fluorescent Antibody Technique
  • HEK293 Cells
  • Humans
  • Immunoglobulin G / blood*
  • Immunoglobulin G / classification
  • Neuromyelitis Optica / blood
  • Neuromyelitis Optica / immunology*
  • Predictive Value of Tests
  • Recombinant Fusion Proteins / immunology
  • Sensitivity and Specificity
  • Transfection

Substances

  • AQP4 protein, human
  • Aquaporin 4
  • Autoantibodies
  • Biomarkers
  • Immunoglobulin G
  • Recombinant Fusion Proteins