Japanese Encephalitis Virus Induce Immuno-Competency in Neural Stem/Progenitor Cells (original) (raw)
Figure 4
Stimulation of T cell proliferation by NSPCs upon JEV infection.
Control and JEV infected NSPCs were dissociated into single cell suspension and treated with mitomycin C. Purified splenic T cells were used as responders and co-cultured with control and infected NSPCs (stimulators) for 72 hours. MTS reagent was used to detect proliferation and absorbance was measured at 490 nm. Experiments were performed in triplicate. The graph represents the stimulator index (SI), which shows that JEV infected NSPCs have a higher stimulator index, i.e. stimulate greater T cell proliferation compared to control NSPCs (A). * indicates p<0.05 compared to control. Purified splenic T cells were labeled with CFSE and then cultured alone or co-cultured with control and JEV infected NSPCs for 72–80 hours. The cells were then stained for anti-CD3ε antibody and analyzed by FACS. The dot plots represent the population of CD3 detected on FL2 channel and CFSE label on FL1 channel. The decrease of CFSE staining is an indication of proliferation and we quantified the percentage of CD3 +ve cells, which have low CFSE label (upper left quadrant of dot plots) (B). JEV infected NSPCs induced T cell proliferation by 2 fold compared to control NSPCs (p<0.01).