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Inhibition of G-protein βγ signaling enhances T cell receptor-stimulated interleukin 2 transcription in CD4+ T helper cells.

Yost EA, Hynes TR, Hartle CM, Ott BJ, Berlot CH - PLoS ONE (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.

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.

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Related in: MedlinePlus

Gβ1 siRNA potentiates TCR-stimulated increases in IL-2 mRNA levels in primary human CD4+ T cells.Box plots (top) and difference plots (bottom) show data from primary human naïve and memory CD4+ T cells isolated from the blood of 30 healthy donors and stimulated for three days with plate-bound anti-CD3 and soluble anti-CD28 in conditions promoting TH1 (A) or TH2 (B) differentiation. IL-2 mRNA levels were determined by qPCR. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.
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pone.0116575.g003: Gβ1 siRNA potentiates TCR-stimulated increases in IL-2 mRNA levels in primary human CD4+ T cells.Box plots (top) and difference plots (bottom) show data from primary human naïve and memory CD4+ T cells isolated from the blood of 30 healthy donors and stimulated for three days with plate-bound anti-CD3 and soluble anti-CD28 in conditions promoting TH1 (A) or TH2 (B) differentiation. IL-2 mRNA levels were determined by qPCR. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Mentions: As in Jurkat cells, Gβ1 siRNA enhanced TCR-stimulated increases in IL-2 mRNA in primary human naïve and memory CD4+ T cells grown for three days in conditions promoting differentiation of TH1 (Fig. 3A) and TH2 (Fig. 3B) T helper cell subsets. Again, despite the fact that TCR stimulation caused larger increases in IL-2 mRNA in TH1 cells compared to TH2 cells and in naïve T cells compared to memory T cells, each of the T cell subsets exhibited significant increases in median TCR-stimulated IL-2 mRNA levels (1.2 to 1.5-fold depending on the subset) in response to Gβ1 siRNA.


Inhibition of G-protein βγ signaling enhances T cell receptor-stimulated interleukin 2 transcription in CD4+ T helper cells.

Yost EA, Hynes TR, Hartle CM, Ott BJ, Berlot CH - PLoS ONE (2015)

Gβ1 siRNA potentiates TCR-stimulated increases in IL-2 mRNA levels in primary human CD4+ T cells.Box plots (top) and difference plots (bottom) show data from primary human naïve and memory CD4+ T cells isolated from the blood of 30 healthy donors and stimulated for three days with plate-bound anti-CD3 and soluble anti-CD28 in conditions promoting TH1 (A) or TH2 (B) differentiation. IL-2 mRNA levels were determined by qPCR. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4309538&req=5

pone.0116575.g003: Gβ1 siRNA potentiates TCR-stimulated increases in IL-2 mRNA levels in primary human CD4+ T cells.Box plots (top) and difference plots (bottom) show data from primary human naïve and memory CD4+ T cells isolated from the blood of 30 healthy donors and stimulated for three days with plate-bound anti-CD3 and soluble anti-CD28 in conditions promoting TH1 (A) or TH2 (B) differentiation. IL-2 mRNA levels were determined by qPCR. **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.
Mentions: As in Jurkat cells, Gβ1 siRNA enhanced TCR-stimulated increases in IL-2 mRNA in primary human naïve and memory CD4+ T cells grown for three days in conditions promoting differentiation of TH1 (Fig. 3A) and TH2 (Fig. 3B) T helper cell subsets. Again, despite the fact that TCR stimulation caused larger increases in IL-2 mRNA in TH1 cells compared to TH2 cells and in naïve T cells compared to memory T cells, each of the T cell subsets exhibited significant increases in median TCR-stimulated IL-2 mRNA levels (1.2 to 1.5-fold depending on the subset) in response to Gβ1 siRNA.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.

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.

Show MeSH
Related in: MedlinePlus