Search

The imidazoquinoline compound R848 is a specific agonist of toll-like receptor (TLR) 7/TLR8 that has been used as an immunostimulant in humans against viral diseases. Although R848-induced immune response has been reported in teleost fish, the relevant mechanism is not clear. In this study, we investigated the antiviral potential and the signaling pathway of R848 in a model of Japanese flounder ( Paralichthys olivaceus ). We found that R848 was able to inhibit the replication of megalocytivirus, stimulated the proliferation of peripheral blood leukocytes (PBL), enhanced the expression of immune genes, and reduced apoptosis of PBL. When endosomal acidification was blocked by chloroquine (CQ), R848-mediated antiviral activity and immune response were significantly reduced. Likewise, inhibition of Myd88 activation markedly impaired the pro-proliferation and anti-apoptosis effect of R848. Cellular study showed that cultured founder cells treated with R848 exhibited augmented NF-κB activity, which, however, was dramatically reduced in the presence of CQ and Myd88 inhibitor. Furthermore, when NF-κB was inactivated, the effect of R848 on cell proliferation and apoptosis was significantly decreased. Taken together, these results indicate that R848 is an immunostimulant with antiviral property in a teleost species, and that the immune response of R848 is mediated by, most likely, TLR7/TLR8 signaling pathway, in which Myd88 and NK-κB play an essential role. [ABSTRACT FROM AUTHOR]

Highlights: [•] Three CpG ODNs that inhibited megalocytivirus replication in flounder were selected. [•] ODN C7 was able to promote leukocyte and phagocyte activation. [•] Knockdown of toll-like receptor 9 expression blocked C7-mediated immune response. [•] C7 exhibited adjuvant property and augmented the protection of a DNA vaccine. [•] C7 enhanced the expression of vaccine-induced genes of innate and adaptive immunity. [ABSTRACT FROM AUTHOR]

Bmal1 is the key component of the central molecular clock that mediates the circadian control of various biological activities including immune reactions. In fish, the role of Bmal1 in immunity is unclear. In this study, we examined the expressional regulation and potential role of Japanese flounder Bmal1 (named PoBmal1) in pathogen associated immune response. PoBmal1 shares 85% identity with its human counterpart and possesses the conserved helix-loop-helix domain and PAS domain. PoBmal1 transcript was distributed extensively in nine tissues and altered to significant extents during the infection of two bacterial pathogens. PoBmal1 , as well as IL-1β, IL-6, IL-8, and TNF-α, exhibited oscillating expression through the light-dark cycle in a tissue-dependent manner, and the expression patterns of the cytokines were largely similar to that of PoBmal1 , both being higher during the photophase. Knockdown of PoBmal1 reduced the expressions of two clock genes (Cry1 and Cry2) and IL-1β, IL-6, IL-8, and TNF-α in a time-dependent manner. During bacterial infection, fish with PoBmal1 knockdown exhibited significantly increased expression of inflammatory cytokines in a tissue- and time-dependent fashion, which largely correlated with significantly decreased bacterial dissemination in tissues. Taken together, our study revealed that flounder Bmal1 exerted a regulatory effect on inflammation response in a manner that was situation-dependent, and that Bmal1 had a significant impact on the outcome of bacterial infection. These results provided new insights into the immune function of fish Bmal1. • PoBmal1 expression occurred in 9 tissues and was regulated by bacterial infection. • PoBmal1 and inflammatory cytokine expressions oscillated similarly with light input. • PoBmal1 knockdown impaired the expressions of Cry and inflammatory cytokines. • PoBmal1 knockdown enhanced inflammatory cytokine expression during bacterial infection. • PoBmal1 knockdown reduced bacterial dissemination in fish tissues. [ABSTRACT FROM AUTHOR]