Evaluation of associated pathways in patients with signs of immune dysregulation by RT-PCR

Uncontrolled activation of immune pathways, so called immune dysregulation may result in autoimmunity, lymphoproliferation, and malignancy. Apoptosis, nuclear factor-κB (NFκB), and phosphoinositide 3-kinase (PI3K) pathways are the major pathways involved in immune dysregulation. The aim of this study was to evaluate the patients with immune dysregulation with selected inflammatory genes' expression analyses. For this reason, some molecules from pathways previously reported to be associated with immune dysregulation in the literature will be selected and their gene expressions will be compared with controls. We evaluated the patients with immune dysregulation, grouped them as those with autoimmune lymphoproliferative syndrome, immune cytopenia, and patients with EBV-associated lymphoproliferation. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was conducted for the selected genes (CASP8, CASP10, FAS, FASL, AKT, MAP3K7, MAP3K14, mTOR, NFKB1, NFKB2, NFKBIA, TRAF3) in inflammatory pathways. The fold changes in gene expression were evaluated using the logarithmic Delta-Delta Ct (2ˆ(–delta delta CT)) method. A statistically significant difference was found comparing the total patient and control groups for relative quantification values of all target genes. No significant difference was observed between the patient groups in terms of target gene expression. According to ROC curve analysis, the target genes with the highest diagnostic rate in terms of distinguishing patients and controls are NFKB2, MAP3K7, AKT, mTOR, NFKB1 and FAS. With this study, we designed an immune dysregulation score/signature by using NFKB1, NFKB2, MAP3K7, AKT, mTOR and FAS for patients suffering from immunological dysregulation, particularly those with immune cytopenia and lymphoproliferation, and can be used in patient diagnosis and follow-up.