Inhibition of Gαs/cAMP Signaling Decreases TCR-Stimulated IL-2 transcription in CD4(+) T Helper Cells.
BACKGROUND
The role of cAMP in regulating T cell activation and function has been controversial. cAMP is generally known as an immunosuppressant, but it is also required for generating optimal immune responses. As the effect of cAMP is likely to depend on its cellular context, the current study investigated whether the mechanism of activation of Gαs and adenylyl cyclase influences their effect on T cell receptor (TCR)-stimulated interleukin-2 (IL-2) mRNA levels.


METHODS
The effect of blocking Gs-coupled receptor (GsPCR)-mediated Gs activation on TCR-stimulated IL-2 mRNA levels in CD4(+) T cells was compared with that of knocking down Gαs expression or inhibiting adenylyl cyclase activity. The effect of knocking down Gαs expression on TCR-stimulated cAMP accumulation was compared with that of blocking GsPCR signaling.


RESULTS
ZM-241385, an antagonist to the Gs-coupled A2A adenosine receptor (A2AR), enhanced TCR-stimulated IL-2 mRNA levels in primary human CD4(+) T helper cells and in Jurkat T cells. A dominant negative Gαs construct, GαsDN3, also enhanced TCR-stimulated IL-2 mRNA levels. Similar to GsPCR antagonists, GαsDN3 blocked GsPCR-dependent activation of both Gαs and Gβγ. In contrast, Gαs siRNA and 2',5'-dideoxyadenosine (ddA), an adenylyl cyclase inhibitor, decreased TCR-stimulated IL-2 mRNA levels. Gαs siRNA, but not GαsDN3, decreased TCR-stimulated cAMP synthesis. Potentiation of IL-2 mRNA levels by ZM-241385 required at least two days of TCR stimulation, and addition of ddA after three days of TCR stimulation enhanced IL-2 mRNA levels.


CONCLUSIONS
GsPCRs play an inhibitory role in the regulation of TCR-stimulated IL-2 mRNA levels whereas Gαs and cAMP can play a stimulatory one. Additionally, TCR-dependent activation of Gαs does not appear to involve GsPCRs. These results suggest that the context of Gαs/cAMP activation and the stage of T cell activation and differentiation determine the effect on TCR-stimulated IL-2 mRNA levels.
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Thomas R
TR
Hynes
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Evan A
EA
Yost
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Stacy M
SM
Yost
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Cassandra M
CM
Hartle
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Braden J
BJ
Ott
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania, 17822-2623, United States of America.
Catherine H
CH
Berlot
G-protein-coupled receptor; Gαs; IL-2; T helper cells; cAMP; heterotrimeric G-protein