Investigation of signaling pathway activations in Multiple Sclerosis of CD3+ t cells

Multiple sclerosis (MS) is an autoimmune disorder prompted by infiltration of auto-reactive immune cells into the central nervous system (CNS). Akt/PKB signaling pathway has an importance in T cell development and survival. Studies revealed the loss of CD4+CD25+CD127-/low Regulatory T (Treg) cell suppression in MS patients. The relationship between the levels of the Akt and phosphorylated-Akt (pAkt) molecule in Treg cells, and the disease severity was investigated. Three different patient groups selected. These are clinically isolated syndrome (CIS, n=8), Benign (n=10) and relapsing remitting multiple sclerosis (RR-MS, n=7). Age and sex matched healthy individuals (n=20) were also involved. Peripheral blood mononuclear cells (PBMC) were seperated from heparinized blood samples by ficoll concentration gradient centrifugation method. Anti-CD3, -CD4, -CD25, -CD127 monoclonal antibodies were used to identify the Treg cells. After stimulus for 15 min with PMA/Ionomycin intracellular staining was made to detect the Akt1 (Ser473) and p-Akt levels in Treg cells. Data were collected and analyzed with FACS DiVa software and FACSAria II with flow cytometer. In the study, the decreases in the level of p-Akt and total Akt1 in MS subjects compared to the healthy groups presents sign that they play a role in progression of disorder. Annual attacks number and progression index of the patients were found to be positively correlated with pAkt and Akt1. The relationship between the Akt molecule and the disease progression was established, and findings were indicate that Akt might be a biomarker for prognosis or therapeutic target molecule.