Ex vivo detection of myelin basic protein-reactive T cells in multiple sclerosis and controls using specific TCR oligonucleotide probes
✍ Scribed by Jian Hong; Ying C. Q. Zang; Sufang Li; Victor M. Rivera; Jingwu Z. Zhang
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 524 KB
- Volume
- 34
- Category
- Article
- ISSN
- 0014-2980
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
T cell reactivity to candidate myelin autoantigens, such as myelin basic protein (MBP), may play an important role in the pathogenesis of multiple sclerosis (MS). Although MBP‐reactive T cellshave been found to undergo in vivo activation in patients with MS, their true precursor frequency in MS is unknown as current frequency analysis is commonly based on the T cell functional responses to MBP. In this study, we developed a TCR sequence‐based ex vivo detection system using colony hybridization with oligonucleotide probes specific for CDR3 of selected T cell clones for the analysis of true T cell precursor frequency in PBMC. The results revealed that the precursor frequency of five independent T cell clones recognizing the immunodominant MBP~83–99~ region was found to be in the range of 1.6×10^–4^ in total T cells in three HLA‐DR2 patients with MS compared to that of 0.25×10^–4^ in HLA‐DR2 healthy individuals. The observed frequency of MBP~83–99~–reactive T cells in MS patients was considerably higher than those measured in parallel by cell culture‐based analysis (2.3×10^–6^) or by enzyme‐linked immunospot assay (3.9×10^–5^) in the same peripheral blood mononuclear cell specimens. Furthermore, the study showed that MBP~83–99~–reactive T cells detected ex vivo belonged to CD45RA^+^, CD25^+^ and CD95^–^ T cell subsets as evidenced by preferential expression of specific TCR transcripts in these cell fractions.