Extended Data Fig. 7: o2R and o9R signalling in T cells engineered with an anti-mesothelin CAR.
From: Potentiating adoptive cell therapy using synthetic IL-9 receptors
a, Schematic of primary mouse CD3+ T cells expressing anti-mesothelin CAR and o2R (CAR-o2R) or o9R (CAR-o9R). b, Representative mesothelin CAR expression (top panel), IL-2Rβ expression (middle panel) and CAR/IL-2Rβ co-expression (bottom panel) in untransduced (UTD) or retrovirally transduced CAR-o2R and CAR-o9R T cells as determined by flow cytometry. Middle panel inset, mean fluorescence intensity (MFI) of IL-2Rβ in untransduced and transduced T cell. c,d, Expansion and phenotype of o2R and o9R CAR T cells. c, In vitro CAR T cell expansion. CAR-o2R and CAR-o9R cells were incubated in the presence of MSA-IL2 (100nM) or MSA-oIL2 (5 μM). An aliquot of cells was removed from the plate and stained with Calcein AM viability dye and counted on the Celigo Image Cytometer daily. Mean ±SEM, n = 3 replicate wells/group. d, CD44 and CD62L co-expression on CAR T cells. Full data of representative flow plots in Fig 3d. CAR-o2R and CAR-o9R cells were incubated for four days in the presence of MSA-IL2 (100nM) or MSA-oIL2 (5 μM). CD44 and CD62L surface co-expression was determined by flow cytometry on live CAR+ cells. Mean ±SEM, n = 3/group. ns, not significant. ****P < 0.0001 (ANOVA). e, Individual growth curves of PDA7940b tumours by treatment group; control groups corresponding to Fig. 3i,j
