MYXV downregulates the expression IL-2, IL2Rα, and IFN-γ in activated human T cells. To determine whether MYXV infection affects the expression of IL-2 and the IL-2α chain receptor (IL-2Rα, aka CD25), about 1 × 10⁶ of mock-treated (ie, without adding virus) or MYXV-treated human T cells and stimulated with anti-CD3/CD28–coated microbeads were culturing for 72 hours, or 96 hours at 37°C, 5% CO₂. Supernatants were collected and analyzed using human IL-2 or human IL-2Rα ELISA. (A-B) MYXV decreases the secretion of IL-2 compared with mock-treated and stimulated T cells. (C-D) Soluble IL-2Rα was significantly downregulated upon infection of activated T cells with MYXV at the indicated time points. (E) Histograms generated from the flow cytometric analysis suggest that MYXV also inhibits the expression of the surface IL-2Rα (CD25) in stimulated and full responder samples (green histograms) as compared with mock-treated and stimulated samples (red histograms). Black histograms correspond to mock-treated T cells and unstimulated T cells, whereas blue histograms correspond to MYXV-treated and unstimulated T cells. (F-G) From the histograms of stimulated samples shown in panel E, the mean fluorescent intensity (MFI) was calculated and is reported as the percentage relative to mock. (F) The MFI of CD25 gated on CD4⁺. (G) The MFI of CD25 gated on CD8⁺. Results represent the mean ± standard error of the mean (SEM) of at least 4 different donors. (H) MYXV did not affect the levels of expression of CD25 in the surface of activated lymphocytes of partial responders. (I-J) MYXV affects the expression of IFN-γ, IL-4, and IL-10 in activated human T cells. After culturing activated T cells for 72 hours or 96 hours, with or without virus infection, 1 × 10⁶ of cells were pelleted and the supernatants of both (I) full responders and (J) partial responders used to evaluate the levels of secreted cytokines such as IL-4, IL-10, and IFN-γ utilizing a Luminex platform. MYXV inhibited the secretion of IFN-γ in all donors tested (I-J) at 72 hours, or 96 hours following stimulation as compared with mock-treated samples; whereas the secretion of the cytokines IL-4 and IL-10 was not affected by MYXV-treated vs mock-treated T cells. Results shown correspond to the mean ± SEM of at least 3 different full responder donors, and 3 different partial responder donors.