Preclinical dose-escalation studies in NHPs. Studies in NHPs (rhesus macaque) injected with transfected autologous apheresis cells. (A) FACS analysis of GFP expression in apheresis products from ZG21 or ZH32 “treated” with GFP mRNA transfection or “untreated.” Side scatter (SSC) by forward scatter (FSC) panel indicates gating for panels to the right. Right panels measure GFP expression in the CD18⁺ granulocyte/monocyte populations (percentage of cells indicated). Baseline GFP-bright clusters are evident in “untreated” panels. (B) FACS analysis of GFP expression in PB from ZG21 or ZH32 “preinfusion” at 1 or 2 days postinfusion of 1 × 10⁸ transfected apheresis cells. SSC by FSC dot plot indicates gating enriched for granulocyte/monocyte population. Two boxed areas in each panel indicate baseline bright GFP⁺ cluster (right box) and less bright GFP⁺ cluster (left box) measuring GFP expression from GFP mRNA transfection. (C) FACS analysis over time after infusion of the less bright GFP cluster (left box value per panel B) in PB from ZG21 (red lines) or ZH32 (blue line) at 5 and 10 minutes, 1 hour, and daily as indicated after infusion of indicated number of autologous transfected apheresis cells.