Function And Mechanism Study Of CAMP Directly Activated Protein Epac-1 In Viral Infection

By the end of 2014,About thirty-six million and nine hundred thousand patients infected with HIV.As a simle 9.2kb-long RNA virus,HIV have successfully avoided the detection of the body's immune system,and ultimately destroy the body's immune defense mechanism.The HIV is overwhelmingly powerful because it took advantage of the host cell immune system and exploit it for the virus's replication and infection.GPCRs and its downstream signaling pathway play a significant role in the pathogenesis of HIV.There has been reported in the journal that,different viruses such as EBv,HCMV,Myxoma virus etc.have the ablity to "Hijack" GPCRs and its downstream signaling pathway regulating the release of cytokines and chemokines,so as to play a role in reducing host immunity and promoting viral infection.This study explored the function and mechanism of GPCRs' downstream cAMP/Epac signaling pathway in antiviral immunity,How virus "deceive" the intracellular signaling pathways for its own good,providing basic research and theoretical support for elucidating its clinical antiviral functionCell line HeLa&Vero,which were originally selected from different tissues and species,were employed in this research.The proliferation of the cell were detected by CKK-8,the expression levels of Epac-1 gene and Vesicular Stomatitis Virus mRNA were assayed through RT-PCR,the amount of VSVG,Epac-1 protein were detected by Western Blot,and the concentrations of cAMP were analyzed by Elisa approach.Finally,all experimental data was statistically analyzed by graphpad.The results showed that the virus could induce the increase of the secondary messenger-cAMP concentration and increase the expression of Epac-1 protein.In return,high concentrations of cAMP in the process of virus infection has played a role in promoting infection.For knockout/inhibition of protein Epac-1 the cAMP downstream effector,it plays a significant immunoprotective effect in viral infection.Also,high expression levels of IL-29 gene were assayed through RT-PCR in Epac-1 Knockout cell line.When the effect of the interleukin 29 was inhibited by the inhibitor Suplatast Tosylate,Epac-1 Knockout cells failed to show its immunoprotective effect in viral infection.These results suggest that the cAMP/EPAC 1 signaling pathway may play an immunomodulatory role by modulating the release of cytokine-interleukin-29.Blocking of cAMP/Epac signaling pathway can cause elevated expression of interleukin 29 in HeLa cells,Interleukin 29 may up-regulate the expression of various immunoreactive factors such as antiviral protein Mx by activating the expression of ISGs,this is an important antiviral effect of immunization pathways.In summary,knockout/inhibition of the cAMP downstream effector protein Epac-1 may play an anti-viral and immunomodulatory role by activating or upregulating the expression of IL-29.