title: Oxysterols are agonist ligands of RORγt and drive Th17 cell differentiation.
abstract: The RAR-related orphan receptor gamma t (RORγt) is a nuclear receptor required for generating IL-17-producing CD4(+) Th17 T cells, which are essential in host defense and may play key pathogenic roles in autoimmune diseases. Oxysterols elicit profound effects on immune and inflammatory responses as well as on cholesterol and lipid metabolism. Here, we describe the identification of several naturally occurring oxysterols as RORγt agonists. The most potent and selective activator for RORγt is 7β, 27-dihydroxycholesterol (7β, 27-OHC). We show that these oxysterols reverse the inhibitory effect of an RORγt antagonist, ursolic acid, in RORγ- or RORγt-dependent cell-based reporter assays. These ligands bind directly to recombinant RORγ ligand binding domain (LBD), promote recruitment of a coactivator peptide, and reduce binding of a corepressor peptide to RORγ LBD. In primary cells, 7β, 27-OHC and 7α, 27-OHC enhance the differentiation of murine and human IL-17-producing Th17 cells in an RORγt-dependent manner. Importantly, we showed that Th17, but not Th1 cells, preferentially produce these two oxysterols. In vivo, administration of 7β, 27-OHC in mice enhanced IL-17 production. Mice deficient in CYP27A1, a key enzyme in generating these oxysterols, showed significant reduction of IL-17-producing cells, including CD4(+) and γδ(+) T cells, similar to the deficiency observed in RORγt knockout mice. Our results reveal a previously unknown mechanism for selected oxysterols as immune modulators and a direct role for CYP27A1 in generating these RORγt agonist ligands, which we propose as RORγt endogenous ligands, driving both innate and adaptive IL-17-dependent immune responses.
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
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authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Jiejun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Xue
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Jiao
authors.initials: J
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authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Steven W
authors.initials: SW
authors.lastname: Sutton
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Marciano
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authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Jingxue
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authors.affiliation: Janssen Research and Development, Spring House, PA 19002.
authors.forename: Marina I
authors.initials: MI
authors.lastname: Nelen
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Liu
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Glenda
authors.initials: G
authors.lastname: Castro
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Rosa
authors.initials: R
authors.lastname: Luna
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Shelby
authors.initials: S
authors.lastname: Crawford
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Homayon
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authors.lastname: Banie
authors.affiliation: Janssen Research and Development, Spring House, PA 19002.
authors.forename: Rose A
authors.initials: RA
authors.lastname: Dandridge
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Xiaohu
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authors.lastname: Deng
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Anton
authors.initials: A
authors.lastname: Bittner
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Chester
authors.initials: C
authors.lastname: Kuei
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Mandana
authors.initials: M
authors.lastname: Tootoonchi
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Natasha
authors.initials: N
authors.lastname: Rozenkrants
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Krystal
authors.initials: K
authors.lastname: Herman
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Jingjin
authors.initials: J
authors.lastname: Gao
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Xia V
authors.initials: XV
authors.lastname: Yang
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Kacey
authors.initials: K
authors.lastname: Sachen
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Karen
authors.initials: K
authors.lastname: Ngo
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Wai-Ping
authors.initials: WP
authors.lastname: Fung-Leung
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Steven
authors.initials: S
authors.lastname: Nguyen
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Aimee
authors.initials: A
authors.lastname: de Leon-Tabaldo
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Blevitt
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Janssen Research and Development, Spring House, PA 19002.
authors.forename: Maxwell D
authors.initials: MD
authors.lastname: Cummings
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Tadimeti
authors.initials: T
authors.lastname: Rao
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Neelakandha S
authors.initials: NS
authors.lastname: Mani
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Changlu
authors.initials: C
authors.lastname: Liu
authors.affiliation: Janssen Research and Development, Spring House, PA 19002.
authors.forename: Murray
authors.initials: M
authors.lastname: McKinnon
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Marcos E
authors.initials: ME
authors.lastname: Milla
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Fourie
authors.affiliation: Janssen Research and Development, LLC, San Diego, CA 92121; and ssun1@its.jnj.com.
authors.forename: Siquan
authors.initials: S
authors.lastname: Sun
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D053493:Cholestanetriol 26-Monooxygenase / Q000378:metabolism; D020381:Interleukin-17 / Q000096:biosynthesis; D008024:Ligands; D051379:Mice; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000819:agonists* / Q000378:metabolism; D013261:Sterols / Q000378:metabolism / Q000494:pharmacology*; D058504:Th17 Cells / Q000166:cytology*
chemical_list: D020381:Interleukin-17; D008024:Ligands; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D013261:Sterols; D053493:Cholestanetriol 26-Monooxygenase; C506832:Cyp27a1 protein, mouse
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