Protocol

Autoimmunity

Volume 102 of the series Methods in Molecular Medicine™ pp 155-171

Evaluation of Autoimmunity to Transaldolase in Multiple Sclerosis

  • Brian NilandAffiliated withDepartments of Medicine, Microbiology, and Immunology, College of Medicine, Upstate Medical University, State University of New York
  • , Andras PerlAffiliated withDivision of Rheumatology, Departments of Medicine and of Microbiology and Immunology, College of Medicine, Upstate Medical University, State University of New York

* Final gross prices may vary according to local VAT.

Get Access

Summary

Transaldolase is a target of autoimmunity mediated by T cells and antibody (Ab) in patients with multiple sclerosis. Functional T-cell assays, T- and B-cell epitope mapping, and detection of transaldolase-specific antibodies in patients with multiple sclerosis are described. Recombinant transaldolase was produced in a prokaryotic expression vector for use in Western blot analysis of sera of these patients. Overlapping transaldolase peptides 15 amino acids (aa) long were synthesized onto cellulose membranes to map immunodominant B-cell epitopes. Amino acid sequence homologies between viral peptides and immunodominant B-cell epitopes of transaldolase were identified using a computer-based algorithm. Direct assessment of molecular mimicry between transaldolase B-cell epitopes and related viral peptides is also shown. T-cell epitopes are mapped in a T-cell proliferation assay using multiple sclerosis patient and control donor cells. Autoantigen-specific T cells are identified by MHC-peptide tetramer staining using flow cytometry analysis.

Key Words

Autoimmunity epitope mapping MHC-tetramer molecular mimicry multiple sclerosis transaldolase