Abstract
Multiple sclerosis (MS) is a chronic inflammatory, demyelinating disease of the central nervous system (CNS), of unknown aetiology and constitutes the main cause of disability in young adults. Autoreactive T lymphocytes against myelin determinants are considered to be implicated in its pathogenesis. T lymphocytes with their antigen receptor, T cell antigen receptor (TCR), recognize antigenic peptides as a complex with MHC molecules. Variable region (V) of α and β‚ chains of TCR is responsible for the specificity of TCR for the antigen. The genes that encode the variable region of TCR ‚ chain are grouped in 25 families. The aim of this study is the determination of expression of TCR V‚ families of TCR in CD4⁺ and CD8⁺ T lymphocytes of peripheral whole blood in patients with multiple sclerosis at the time of diagnosis and in patients with “relapsing-remitting” form of the disease. For the conduction of the study, totally 84 individuals were examined during March 2003 and September 2006. ...
Multiple sclerosis (MS) is a chronic inflammatory, demyelinating disease of the central nervous system (CNS), of unknown aetiology and constitutes the main cause of disability in young adults. Autoreactive T lymphocytes against myelin determinants are considered to be implicated in its pathogenesis. T lymphocytes with their antigen receptor, T cell antigen receptor (TCR), recognize antigenic peptides as a complex with MHC molecules. Variable region (V) of α and β‚ chains of TCR is responsible for the specificity of TCR for the antigen. The genes that encode the variable region of TCR ‚ chain are grouped in 25 families. The aim of this study is the determination of expression of TCR V‚ families of TCR in CD4⁺ and CD8⁺ T lymphocytes of peripheral whole blood in patients with multiple sclerosis at the time of diagnosis and in patients with “relapsing-remitting” form of the disease. For the conduction of the study, totally 84 individuals were examined during March 2003 and September 2006. Fifty two of them were patients with multiple sclerosis, with varying degree of disability, from outpatient clinic for multiple sclerosis of 2nd Neurological clinic of AHEPA Hospital. Twenty of them, the first group, were patients at the time of diagnosis of multiple sclerosis, aged between 14 to 53 years old (mean age: 35,2 years). Thirty two of them, the second group, were patients with multiple sclerosis of “relapsing remitting” form of the disease aged between 25 to 56 years old (mean age: 40,2 years). The sample also included 32 age-matched, healthy adults that were used as control group. One sample was taken from each individual containing peripheral whole blood with anticoagulant EDTA. Specifically for the first group, three samples were taken, one initially and two others during the follow-up period. With the use of flow cytometry of three fluorescence, the proportion of expression of families of ‚ chain’s variable region of T cell antigen receptor (TCR V‚) was determined. In certain samples the molecular technique RT-PCR was also used to analyse the recombination of TCR V‚ genes. The results showed the following: Statistically significant alteration of certain TCR V‚ families was observed between patients of the first group and healthy controls, concerning CD8⁺ T lymphocytes. In this group was observed oligoclonal expansion of one TCR V‚ family, concerning also CD8⁺ T lymphocytes. During the follow-up period, increase was observed in some patients and in others reduction of the proportion of expression of individual TCR V‚ families. In patients of the second group, compared to healthy individuals, was observed statistically significant alteration of less TCR V‚ families than the first group. Oligoclonal expansions concerning also CD8⁺ T lymphocytes were observed in patients of this group in a higher percentage compared to healthy individuals. The TCR V‚ family expansions were individual-specific (private) for each patient. TCR V‚ family expansion was not found to characterise neither multiple sclerosis nor each individual group of the disease. Monoclonal expansion of TCR V‚ family was not observed in any patient. Our study notified the importance of TCR in the pathogenesis of multiple sclerosis, suggesting that blood T cell repertoire analysis could be used as an additional non invasive method for disease follow-up in Multiple Sclerosis. Finally, flow cytometry appeared to be a useful applicable method for this follow-up in daily practice with the advantage of quantitative determination of TCR V‚ family expression.
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