Project description:We used the BDA DNBelabC4 platform to conduct single-cell RNA transcriptome sequencing on hemocytes before and after infection with WSSV, and to study the changes in hemocytes composition and antiviral mechanism of a crustacean virus infection represented by mud crab. The blood cells, or hemocytes, of the mud crab (Scylla paramamosain) play crucial roles in its innate immune system. These cells are involved in various immune responses, including phagocytosis, encapsulation, and the production of antimicrobial peptides. The hemocytes can be isolated and characterized using techniques such as flow cytometry, which helps in understanding their functions and interactions within the crab's immune system. White Spot Syndrome Virus (WSSV) is a highly contagious and lethal virus that primarily infects crustaceans, particularly shrimp and crabs. It is an enveloped double-stranded DNA virus, typically oval-shaped, measuring about 320nm in length and 100nm in width, making it one of the largest known viruses. WSSV is the causative agent of White Spot Syndrome, a disease that has caused significant economic losses in the aquaculture industry.
Project description:The phenomenon of trained immunity, which facilitates vaccine development for disease control, has been identified in shrimp; however, the mechanism remains elusive. In the present study, we found that histone H3K27 acetylation (H3K27ac) mediated by the lysine acetyltransferase KAT8 plays an important role in preventing white spot syndrome virus (WSSV) infection in the shrimp Marsupenaeus japonicus. We then successfully established a model of trained immunity via the use of UV-inactivated WSSV to explore the underlying mechanism(s) in shrimp. In UV-WSSV-trained shrimp, the glycolysis and tricarboxylic acid (TCA) cycle metabolic pathways were enhanced and acetyl-CoA concentrations were increased. As the acetyl group donor, acetyl-CoA promotes KAT8 activity to increase H3K27 acetylation. H3K27ac is deposited at the promoter region of the transcription factor Dorsal to facilitate its expression and then Dorsal promotes the expression of an interferon-like cytokine, Vago5, and antimicrobial peptides that act against WSSV infection. H3K27ac is also deposited at the promoter region of hexokinase 2 and isocitrate dehydrogenase, which positively regulates glycolysis and the TCA cycle in a feedforward manner. Our results reveal a novel mechanism of trained immunity induced by UV-WSSV in shrimp and provide a theoretical basis for the development of antiviral vaccines for disease control in shrimp aquaculture.
Project description:In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis, the expressions of viral miRNAs were tissue-specific in vivo. In this study, the viral miRNAs from white spot syndrome virus (WSSV) were characterized in shrimp in vivo. On the basis of our previous study and small RNA sequencing in this study, a total of 89 putative WSSV miRNAs were identified. As revealed by miRNA microarray analysis and Northern blots, the expressions of viral miRNAs were tissue-specific in vivo. Therefore, our study presented the first report on the in vivo molecular events of viral miRNA in the antiviral apoptosis.