Fig. 2

Compound 9 reverses arginine depletion mediated interferon gamma production. a Arginine depletion prevented IFNg secretion by splenocytes. Splenocytes were activated with CD3/28 in the presence of media with arginine levels of 0, 2.5, 10, 40, 160 and 1000 uM. After 24 h IFNg was determined by ELISA in the culture supernatants b-d. Compound 9 mediated restoration of T cell function. Splenocytes were isolated from mixed background mice, dissociated and activated with CD3/28 antibodies in the presence of in the presence of arginine 40 uM. Recombinant Arginase (ARG1, 1 μg/ml) and Cpd9 (50 uM) were added to the indicated wells at the same time. b IFNg was determined by ELISA in the culture supernatants after 24 h c Granzyme B expression CD8 T cells were determined by flow cytometry d IL-2 expressing CD8 T cells were determined by flow cytometry. *p < =0.05, **p < =0.01, and ***p < =0.001. Graphs show representative results from experiments performed at least three times