Defining the AHR-regulated transcriptome in NK cells reveal gene expression programs relevant to development and function

Aryl hydrocarbon receptor (AHR) is a ligand activated transcription factor that plays a critical role in shaping innate and adaptive immune responses. However, the molecular mechanism by which AHR regulates the function of human NK cells remains largely unknown. Here, we use RNA sequencing on IL21-expanded NK cells in the presence of AHR agonists and antagonist to demonstrate that AHR directly regulates the expression of known regulators of phenotype, development, metabolism, and function. Overall design: NK cells were isolated from leukopak of healthy volunteer donors using NK cell RosetteSep. NK cells were expanded using irradiated feeder cells overexpressing membrane bound IL21 (FC21) for period of 14-21 days. NK cells from 3-donors (NK1, NK124 and NK126) were expanded using human interleukin-2 (IL2-50 IU/ml) alone, IL2+stemregenin (SR1-1uM) or IL2+kynurenine (Kyn-25uM). The media was changed every 2-3 days. The samples for RNA were collected on days 0, 7, 14 and 21 (D0, D7, D14 and D21). RNA was isolated using RNeasy kit all the samples were DNase treated and concentrated using RNA concentrator kit. We compared gene expression profiling between IL2 vs SR1 and IL2 vs Kyn on days 7 and 14.