title: Natural agonists for aryl hydrocarbon receptor in culture medium are essential for optimal differentiation of Th17 T cells.
abstract: Th17 cell differentiation is dependent on interleukin (IL)-6 and transforming growth factor (TGF)-beta, and it is modulated by activation of the aryl hydrocarbon receptor (AhR). In this study, we show that differentiation of Th17 cells, but not Th1 or induced regulatory T (iT reg) cells, is increased by endogenous AhR agonists present in culture medium. Th17 development from wild-type mice is suboptimal in the presence of the AhR antagonist CH-223191, similar to the situation in AhR-deficient mice, which show attenuated IL-17 production and no IL-22 production. The presence of natural AhR agonists in culture medium is also revealed by the induction of CYP1A1, a downstream target of AhR activation. However, the most commonly used medium, RPMI, supports very low levels of Th17 polarization, whereas Iscove's modified Dulbecco's medium, a medium richer in aromatic amino acids, which give rise to AhR agonists, consistently results in higher Th17 expansion in both mouse and human cells. The relative paucity of AhR agonists in RPMI medium, coupled with the presence of factors conducive to IL-2 activation and enhanced Stat5 phosphorylation, conspire against optimal Th17 differentiation. Our data emphasize that AhR activation plays an essential part in the development of Th17 cells and provide a rational explanation for the poor in vitro polarization of Th17 cells that is reported in the majority of publications for both mouse and human cells.
authors.affiliation: Division of Molecular Immunology, Medical Research Council National Institute for Medical Research, London NW7 1AA, England, UK.
authors.forename: Marc
authors.initials: M
authors.lastname: Veldhoen
authors.forename: Keiji
authors.initials: K
authors.lastname: Hirota
authors.forename: Jillian
authors.initials: J
authors.lastname: Christensen
authors.forename: Anne
authors.initials: A
authors.lastname: O'Garra
authors.forename: Brigitta
authors.initials: B
authors.lastname: Stockinger
mesh_terms: D000818:Animals; D001391:Azo Compounds / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020381:Interleukin-17 / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D010766:Phosphorylation / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists / Q000037:antagonists & inhibitors / Q000502:physiology*; D018168:Receptors, Retinoic Acid / Q000235:genetics; D011988:Receptors, Thyroid Hormone / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050799:STAT5 Transcription Factor / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014364:Tryptophan / Q000494:pharmacology; D000097043:Interleukin-22
chemical_list: D001391:Azo Compounds; D002227:Carbazoles; D003470:Culture Media; D019363:Cytochrome P-450 CYP1A1; D020381:Interleukin-17; D007376:Interleukin-2; D007378:Interleukins; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D011720:Pyrazoles; D018336:Receptors, Aryl Hydrocarbon; D018168:Receptors, Retinoic Acid; D011988:Receptors, Thyroid Hormone; D050799:STAT5 Transcription Factor; D014364:Tryptophan; D000097043:Interleukin-22; C511621:2-methyl-2H-pyrazole-3-carboxylic acid (2-methyl-4-o-tolylazophenyl)amide; C111855:6-formylindolo(3,2-b)carbazole; C539900:RORC protein, human; C539901:Rorc protein, mouse