Evolutionary analyses reveal immune cell receptor GPR84 as a conserved receptor for bacteria-derived molecules

Function and regulation of GPR84 in human neutrophils

“…Interestingly, a recent study showed the bacterial quorum sensing molecules cis-2-decenoic acid and trans-2-decenoic acid were agonists for mammalian GPR84 orthologues (Schulze et al, 2022), but whether these molecules also activate the GPR84 expressed by human neutrophils remains to be determined. To overcome the weak activity of the MCFAs, recent research using GPR84-targeting synthetic tool compounds such as embelin (Gaidarov et al, 2018;Mahmud et al, 2017), 6-n-octylaminouracil (6-OAU [Luscombe et al, 2020Suzuki et al, 2013]) and 2-(hexylthio)pyrimidine-4,6-diol (ZQ16 [Zhang et al, 2016]) has increased our knowledge of GPR84-induced neutrophil functions.…”

“…However, because high concentrations of MCFAs are needed to activate rather modest GPR84‐mediated neutrophil responses in vitro and that it is unclear if such high concentrations are present in vivo, it remains to be elucidated if there are other (more potent) endogenous ligand/ligands for GPR84 (Luscombe et al, 2020). Interestingly, a recent study showed the bacterial quorum sensing molecules cis ‐2‐decenoic acid and trans ‐2‐decenoic acid were agonists for mammalian GPR84 orthologues (Schulze et al, 2022), but whether these molecules also activate the GPR84 expressed by human neutrophils remains to be determined. To overcome the weak activity of the MCFAs, recent research using GPR84‐targeting synthetic tool compounds such as embelin (Gaidarov et al, 2018; Mahmud et al, 2017), 6‐ n ‐octylaminouracil (6‐OAU [Luscombe et al, 2020; Suzuki et al, 2013]) and 2‐(hexylthio)pyrimidine‐4,6‐diol (ZQ16 [Zhang et al, 2016]) has increased our knowledge of GPR84‐induced neutrophil functions.…”

Protocol to characterize Gi/o and Gs protein-coupled receptors in transiently transfected cells using ELISA and cAMP measurements

“… 1 acgtccccgactacgccTGGAACAGCTCTGACGCC FLAG-adapter primer-AS (human GPR84) Schulze et al. 1 TCActtatcgtcatcgtccttatagtcATGGAGCCTATGGAA HA-universal primer EcoRI -S Schulze et al. 1 cgcGAATTCgccaccATGtacccctacgacgtccccgactacg FLAG-universal primer SpeI -AS Schulze et al.…”

“… 1 cgccgcACTAGTTCActtatcgtcatcgtccttatagtc Chemicals, peptides, and recombinant proteins 10× HBSS Thermo Fisher Scientific 14065056 1× D-PBS Thermo Fisher Scientific 14190144 3-Isobutyl-1-methylxanthine (IBMX) Sigma-Aldrich I5879 Bovine serum albumin (BSA) Sigma-Aldrich A3294 Citric acid Sigma-Aldrich 251275 Deoxynucleotide (dNTP) Solution Set NEB N0446 Dulbecco’s Modified Eagle Medium (DMEM) Thermo Fisher Scientific 41966029 Dulbecco’s Modified Eagle Medium (DMEM)/ F-12 Thermo Fisher Scientific 21041025 Dithiothreitol (DTT) Sigma-Aldrich DTT-RO UltraPure 0.5 M EDTA, pH 8 Thermo Fisher Scientific 15575020 Fetal bovine serum (FBS) Thermo Fisher Scientific 10270106 Formaldehyde Sigma-Aldrich F8775 Forskolin Sigma-Aldrich F3917 H 2 O 2 Sigma-Aldrich 1.08600 HCl Sigma-Aldrich 258148 HEPES (1 M) Thermo Fisher Scientific 15630056 Lipofectamine 2000 Thermo Fisher Scientific 11668019 Na 2 HPO 4 · 2 H 2 O Sigma-Aldrich S9763 Na 2 SO 3 Sigma-Aldrich S0505 Nonidet P40 (substitute) (NP-40) Santa Cruz sc-29102 o-phenylenediamine Sigma-Aldrich P23938 Opti-MEM Thermo Fisher Scientific 31985070 ...…”

“…Previously published data following this protocol includes data for hydroxycarboxylic acid receptors and GPR84, 1 , 2 , 3 trace amine-associated receptors 4 , 5 , 6 and melanocortin receptors. 7 , 8 …”