Autoimmunity pp 155-171
Evaluation of Autoimmunity to Transaldolase in Multiple Sclerosis
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 WordsAutoimmunity epitope mapping MHC-tetramer molecular mimicry multiple sclerosis transaldolase
- 6.Lees, M. B. and Brostoff, S. W. (1984) Proteins of myelin, in Myelin (Morell, P., ed), Plenum Press, New York, pp. 197–224.Google Scholar
- 10.Parker, K. HLA peptide binding predictions. NIH BIMAS website: http://bimas.dcrt.nih.gov/molbio/hla_bind/. Accessed April, 2003.
- 20.Fleiss, J. L. (1981) Statistical Methods for Rates and Proportions, Wiley, New York.Google Scholar
- 27.Kuroda, M. J., Schmitz, J. E., Barouch, D. H., Craiu, A., Allen, T. M., Sette, A., et al. (1998) Analysis of Gag-specific cytotoxic T lymphocytes in simian immunodeficiency virus-infected rhesus monkeys by cell staining with a tetrameric major histocompatibility complex class I-peptide complex. J. Exp. Med. 187, 1373–1381.PubMedCrossRefGoogle Scholar
- 28.National Institutes of Allergy and Infectious Diseases (NIAID), Tetramer Facility Guide, McKesson Biosciences Corp. http://www.niaid.nih.gov/reposit/tetramer/index.html. April 2003