title: Inhibition of G-protein βγ signaling enhances T cell receptor-stimulated interleukin 2 transcription in CD4+ T helper cells.
abstract: G-protein-coupled receptor (GPCR) signaling modulates the expression of cytokines that are drug targets for immune disorders. However, although GPCRs are common targets for other diseases, there are few GPCR-based pharmaceuticals for inflammation. The purpose of this study was to determine whether targeting G-protein βγ (Gβγ) complexes could provide a useful new approach for modulating interleukin 2 (IL-2) levels in CD4+ T helper cells. Gallein, a small molecule inhibitor of Gβγ, increased levels of T cell receptor (TCR)-stimulated IL-2 mRNA in primary human naïve and memory CD4+ T helper cells and in Jurkat human CD4+ leukemia T cells. Gβ1 and Gβ2 mRNA accounted for >99% of Gβ mRNA, and small interfering RNA (siRNA)-mediated silencing of Gβ1 but not Gβ2 enhanced TCR-stimulated IL-2 mRNA increases. Blocking Gβγ enhanced TCR-stimulated increases in IL-2 transcription without affecting IL-2 mRNA stability. Blocking Gβγ also enhanced TCR-stimulated increases in nuclear localization of nuclear factor of activated T cells 1 (NFAT1), NFAT transcriptional activity, and levels of intracellular Ca2+. Potentiation of IL-2 transcription required continuous Gβγ inhibition during at least two days of TCR stimulation, suggesting that induction or repression of additional signaling proteins during T cell activation and differentiation might be involved. The potentiation of TCR-stimulated IL-2 transcription that results from blocking Gβγ in CD4+ T helper cells could have applications for autoimmune diseases.
authors: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
authors: Evan A
authors: EA
authors: Yost
authors: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
authors: Thomas R
authors: TR
authors: Hynes
authors: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
authors: Cassandra M
authors: CM
authors: Hartle
authors: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
authors: Braden J
authors: BJ
authors: Ott
authors: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
authors: Catherine H
authors: CH
authors: Berlot
mesh_terms: D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D044387:GTP-Binding Protein beta Subunits / Q000037:antagonists & inhibitors / Q000378:metabolism*; D044388:GTP-Binding Protein gamma Subunits / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics*; D050778:NFATC Transcription Factors / Q000378:metabolism; D011401:Promoter Regions, Genetic; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D006377:T-Lymphocytes, Helper-Inducer / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors; D014158:Transcription, Genetic; D014966:Xanthenes / Q000494:pharmacology
chemical_list: C118981:G-protein Beta gamma; D044387:GTP-Binding Protein beta Subunits; D044388:GTP-Binding Protein gamma Subunits; D007376:Interleukin-2; D050778:NFATC Transcription Factors; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011948:Receptors, Antigen, T-Cell; D014966:Xanthenes; C007820:gallein; D002118:Calcium
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