Fig. 3

Functional traits of mutein TNF07 compared to sTNF and a TNFR2 antagonistic antibody in CD4 T-regulatory expansion assays. a Flow cytometry pictures of normal CD4 T cells treated for 16 h with IL-2, IL-2 plus sTNF, IL-2 plus TNF07 or IL-2 plus a TNFR2 antagonistic antibody. In this prototype experiment, isolated CD4 T cells with IL-2 incubation alone had a basal level of human T regulatory T cells (CD4 + CD25 + Foxp3+) of 5.2 % after the overnight incubation. In contract, treated with sTNF expanded the Tregs to 6.9 % and TNF07 expanded the Tregs more successfully to 7.1 %. In contrast, the same cells expanded with IL-2 plus a TNFR2 antibody antagonist showed halted Treg expansion (5.3 %). b The experiment is (a) was performed a total of 6 times on different CD4 human donors. Data is represented as the % increase or decrease in Treg cells with sTNF, TNF07 or with the TNFR2 antagonistic antibody compared to IL-2 alone overnight incubations. A = IL2 vs IL2 + Antagonist Ab (p < 0.001), B = IL2 vs IL2 + TNF-07 (p = 0.0003), C = IL2 vs IL2 + sTNF (p = 0.0027). c An inhibition experiment shows the ability of a monoclonal TNFR2 antagonistic antibody with dose escalations (ug/ml) to prevent the agonism of the TNF07 mutein