Natural Killer cells (NKs) are of interest for their ability to modulate immune responses and eliminate infected cells. As a result of these mechanisms, NKs possess the potential to be developed into new immunotherapy protocols. Along with their cytotoxic function, NKs produce immunomodulatory cytokines like IFN-γ upon cytokine or activating receptor stimulation. Our lab is interested in IFN-γ regulation, which is subject to differential transcriptional and translational pathways to IFN-γ protein production based on the activating signal and prior culture conditions. Published research and our preliminary studies suggest transcription factor c-Myc may be involved in Ifng transcription. We hypothesized that Myc acts as a transcription factor for Ifng in high dose IL-15 cultured activating receptor-stimulated NKs. Myc EGFP reporter mice were employed to measure the expression of Myc via flow cytometry, and the mean fluorescence intensity of EGFP was quantified. Acute pharmacological inhibition of Myc and a genetic model of tamoxifen-inducible Myc deletion were pursued. NKs were either freshly isolated, cultured for 72 hours with 10 ng/mL (LD) or with 100ng/mL (HD) IL-15, followed by NK1.1 activating receptor stimulation. Ifng transcript was quantified by RT-PCR and IFN-γ was measured via intracellular flow cytometry. Results were analyzed by a paired, non-parametric t-test. Myc inhibition during activating receptor stimulation of high dose IL-15 cultured NKs showed diminished Ifng transcript. However, the genetic model of Myc deletion did not result in similar changes. IFN-γ protein levels remained unaffected by either.

Dr. Megan Cooper and PhD student Maria Cimpean have been influential parts of Nathan's mentors at WashU. The Cooper lab has pushed him to think critically, answered his many questions, and taught him to love the challenge of science and academic research. Nathan is grateful for the researcher he has developed into through his lab's expertise, encouragement, and support.