Figure 2

Expansion and Profile of nine expanded Treg products: (A) Outline of clinical good manufacturing expansion protocol established at the Mathews Center for Cellular Therapy, Northwestern Memorial Hospital. (B) Absolute cell number of all nine Treg products throughout expansion protocol; black bar represents grand median of all products (n=9). (C) Representative clinical phenotyping of Treg products, specifically CD4⁺CD25⁺FOXP3⁺ expression on post CD25 enrichment (day 0) as well as days 14 and 21 of culture. (D) Represents the average (±SD) of expression of Treg markers CD4; CD25; and FOXP3; also non-Treg markers CD8, CD20, and CD127 throughout expansion protocol (n=9). (E) Heat map of clinical Treg products and controls (day 0) depicting the percentage of methylation within 9 CpG sites of the conserved non-coding sequence 2 of FOXP3 gene in the expanded Tregs (TRK- 01–09; n=4). (F) Average (+SD) of the percent methylation in day 0 control products and day 21 expanded Treg products (n=4). ***p < 0.005, *p < 0.05; GM  =  growth medium (described under Materials).