Shift in CD4
PROLIFERATION, BLOOD
The effect of the Ad5-IL-12 gene therapy on immune system activation was evaluated by analyzing circulating PBMCs by flow cytometry. Patient blood was obtained at least once before the Ad5-IL-12 gene therapy administration (baseline) and on follow-up days as described in the clinical protocol. Blood was processed on the day of draw and collected PBMC’s were frozen at -80C until all the samples were collected from a patient. Subsequently, frozen PBMCs were thawed, processed and quantitated by multi-color flow cytometry as described in Materials and Methods. Antibodies against CD3, CD56, CD4, and CD8 served to measure NK, Th and Tc cells while activation of the immune system was monitored by staining for CD45RO, CD69, and proliferation by Ki67. Antibodies against Tim3 were used to detect suppression of the immune system following treatment [