Fas- and DR3-dependent transcriptome changes in naïve T-cells from EBV carrier during and after acute mononucleosis

EBV-encoded proteins interfere with extrinsic and intrinsic apoptotic pathways so that homeostasis of infected cells shifts to survival and proliferation favoring to evasion of virus from immune surveillance.The aim of investigation is to reveal the key factors regulating Fas- и DR3-triggered cascades influenced by EBV in naïve T-cells. We developed the splicing-sensitive microarray covering 221 genes encoding apoptosis-associated factors, adaptor proteins, kinases, RNA-binding proteins (including spliceosome elements) etc. Naïve CD4+ and CD8+ T-cells isolated from the same EBV-infected patient during acute mononucleosis and after therapy were analyzed. Each microarray encompassed 4 identical probe-containing areas. Each area contains 1042 target probes in PM- and MM-version (2084 total) and 140 negative control probes. MM-probes were excluded from processed data. Freshly isolated naïve T-cells were either subjected to immediate NA extraction or were cultivated for 24 hours in 1) empty media [control]; 2) Fas-agonist containing media [Fas]; 3) DR3-agonist containing media [DR3]. The following comparisons were made: [control]/[Freshly isolated]; [Fas]/[control]; [DR3]/[control]. Experiments were performed for CD4+ and CD8+ T-cells isolated before and after therapy. Additional comparisons [Freshly isolated after]/[Freshly isolated before] were made for each subpopulation. Please note that the data was normalized only within COMPARED samples (like 'Cd4_control_before*' samples vs.'cd4_fas_before*' samples), *not* across ALL the samples.