Melatonin induces nitric oxide and the potential mechanisms relate to innate immunity against bacterial pathogen infection in Arabidopsis

Melatonin (N‐acetyl‐5‐methoxytryptamine) is a naturally occurring small molecule, serving as important secondary messenger in the response of plants to various biotic and abiotic stresses. However, the interactions between melatonin and other important molecules in the plant stress response, especially in plant immunity, are largely unknown. In this study, we found that both melatonin and nitric oxide (NO) levels in Arabidopsis leaves were significantly induced by bacterial pathogen (Pst DC3000) infection. The elevated NO production was caused by melatonin as melatonin application enhanced endogenous NO level with great efficacy. Moreover, both melatonin and NO conferred improved disease resistance against Pseudomonas syringe pv. tomato (Pst) DC3000 infection in Arabidopsis. NO scavenger significantly suppressed the rise of NO which was induced by exogenous application of melatonin. As a result, the beneficial effects of melatonin on the expression of salicylic acid (SA)‐related genes and disease resistance against bacterial pathogen infection were jeopardized by use of a NO scavenger. Consistently, melatonin application significantly lost its effect on the innate immunity against P. syringe pv. tomato (Pst) DC3000 infection in NO‐deficient mutants of Arabidopsis. The results indicate that melatonin‐induced NO production is responsible for innate immunity response of Arabidopsis against Pst DC3000 infection.     

Suppression of adenoviral gene expression in the liver: role of innate vs adaptive immunity and their cell lysis mechanisms.

BACKGROUND: Injection of adenoviral constructs causes liver infection prompting immunity, which suppress viral gene expression. Innate and adaptive immunity mediate these processes raising the question which pathways are the most prominent. METHODS: Adenovirus expressing the beta-galactosidase (beta-gal) gene was injected into normal and immunodeficient mice. Elimination of beta-gal-expressing hepatocytes and increases in liver enzymes were assayed. Major histocompatibility complex (MHC) class I densities, perforin channel insertion and apoptosis by Fas and tumor necrosis factor (TNF)-alpha were assayed. RESULTS: At high virus doses, suppression of viral gene expression was as efficient in immunodeficient as in normal mice, while at low doses effects of cytotoxic T lymphocytes (CTL) were demonstrable. Despite CTL priming and elimination of infected hepatocytes no liver injury is detected. Hepatocyte MHC I densities were able to trigger CTL granule exocytosis and perforin lysis in vitro but not in vivo. This is we show is because of decreased sensitivity of hepatocytes from infected mice to perforin and increased sensitivity to Fas and TNF-alpha lysis. CONCLUSION: Effector cells of the innate immune system are exceedingly effective in suppressing adenoviral gene expression. Perforin-independent pathways, those mediated by TNF-alpha and Fas are very efficient in hepatocytes from virus-infected livers.     

Regulation of microRNA by hepatitis B virus infection and their possible association with control of innate immunity

Hepatitis B virus (HBV) chronically infects more than 350 million people worldwide. HBV causes acute and chronic hepatitis, and is one of the major causes of cirrhosis and hepatocellular carcinoma. There exist complex interactions between HBV and the immune system including adaptive and innate immunity. Toll-like receptors (TLRs) and TLR-signaling pathways are important parts of the innate immune response in HBV infections. It is well known that TLR-ligands could suppress HBV replication and that TLRs play important roles in anti-viral defense. Previous immunological studies demonstrated that HBV e antigen (HBeAg) is more efficient at eliciting T-cell tolerance, including production of specific cytokines IL-2 and interferon gamma, than HBV core antigen. HBeAg downregulates cytokine production in hepatocytes by the inhibition of MAPK or NF-κB activation through the interaction with receptor-interacting serine/threonine protein kinase. MicroRNAs (miRNAs) are also able to regulate various biological processes such as the innate immune response. When the expressions of approximately 1000 miRNAs were compared between human hepatoma cells HepG2 and HepG2.2.15, which could produce HBV virion that infects chimpanzees, using real-time RT-PCR, we observed several different expression levels in miRNAs related to TLRs. Although we and others have shown that HBV modulates the host immune response, several of the miRNAs seem to be involved in the TLR signaling pathways. The possibility that alteration of these miRNAs during HBV infection might play a critical role in innate immunity against HBV infection should be considered. This article is intended to comprehensively review the association between HBV and innate immunity, and to discuss the role of miRNAs in the innate immune response to HBV infection.     

Melatonin is required for H2O2‐ and NO‐mediated defense signaling through MAPKKK3 and OXI1 in Arabidopsis thaliana

Melatonin influences plant innate immunity through the mitogen‐activated protein kinase (MAPK) pathway. However, the most upstream MAPK component in melatonin signaling and the dependence of generation of a reactive oxygen species (ROS) burst on melatonin synthesis and signaling remain unclear. In this study, treatment of several mekk (alias mapkkk)‐knockout Arabidopsis mutants with melatonin revealed that the MAPKKK3 and OXI1 (oxidative signal‐inducible1) kinases are responsible for triggering melatonin‐induced defense signaling pathways. In addition, melatonin induction upon infection with the avirulent pathogen Pseudomonas syringae DC3000 (avrRpt2) was independent of H₂O₂ and NO individually, but dependent on the combination of H₂O₂ and NO. Moreover, melatonin‐mediated induction of the expression of defense‐related genes, such as PR1 and ICS1, was not altered in the H₂O₂‐deficient rbohD/F‐knockout mutant cotreated with an NO scavenger, indicating that melatonin functions downstream of the ROS and NO burst. Collectively, the data indicate that melatonin‐mediated induction of an innate immune response requires multiple signaling molecules and activation of MAPKKK3 and OXI1, followed by triggering of downstream MAPK cascades, such as MAPK3 and MAPK6.