PT - JOURNAL ARTICLE AU - Joseph W. Zagorski AU - Tyler P. Maser AU - Karen T. Liby AU - Cheryl E. Rockwell TI - Nrf2-Dependent and -Independent Effects of <em>tert</em>-Butylhydroquinone, CDDO-Im, and H<sub>2</sub>O<sub>2</sub> in Human Jurkat T Cells as Determined by CRISPR/Cas9 Gene Editing AID - 10.1124/jpet.116.238899 DP - 2017 May 01 TA - Journal of Pharmacology and Experimental Therapeutics PG - 259--267 VI - 361 IP - 2 4099 - http://jpet.aspetjournals.org/content/361/2/259.short 4100 - http://jpet.aspetjournals.org/content/361/2/259.full SO - J Pharmacol Exp Ther2017 May 01; 361 AB - Nuclear factor erythroid 2-related factor 2 (Nrf2) is a stress-activated transcription factor activated by stimuli such as electrophilic compounds and other reactive xenobiotics. Previously, we have shown that the commonly used food additive and Nrf2 activator tert-butylhydroquinone (tBHQ) suppresses interleukin-2 (IL-2) production, CD25 expression, and NFκB activity in human Jurkat T cells. The purpose of the current studies was to determine whether these effects were dependent upon Nrf2 by developing a human Nrf2-null T cell model using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 technology. The current studies show that suppression of CD25 expression by tBHQ is partially dependent on Nrf2, whereas inhibition of IL-2 secretion is largely Nrf2-independent. Interestingly, tBHQ inhibited NFκB activation in an Nrf2-independent manner. This was an unexpected finding since Nrf2 inhibits NFκB activation in other models. These results led us to investigate another more potent Nrf2 activator, the synthetic triterpenoid 1[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl]imidazole (CDDO-Im). Treatment of wild-type and Nrf2-null Jurkat T cells with CDDO-Im resulted in an Nrf2-dependent suppression of IL-2. Furthermore, susceptibility to reactive oxygen species was significantly enhanced in the Nrf2-null clones as determined by decreased mitochondrial membrane potential and cell viability. Importantly, this study is the first to describe the generation of a human Nrf2-null model, which is likely to have multiple applications in immunology and cancer biology. Collectively, this study demonstrates a role for Nrf2 in the effects of CDDO-Im on CD25 and IL-2 expression, whereas the effect of tBHQ on these parameters is complex and likely involves modulation of multiple stress-activated transcription factors, including NFκB and Nrf2.