Molecular characterization of a novel white spot syndrome virus response protein (dubbed LvWRP) from Litopenaeus vannamei.
White spot syndrome, which is caused by white spot syndrome virus (WSSV), is a highly contagious disease of penaeid shrimp. However, there is currently incomplete understanding of the infection mechanism and pathogenesis of WSSV. In this study, a novel gene of a previously uncharacterized WSSV response protein (LvWRP) in Litopenaeus vannamei was identified and characterized. The LvWRP gene has an open reading frame (ORF) of 879 bp encoding a putative protein of 292 amino acids. Sequence analysis revealed that LvWRP shared 24.9% identity with an uncharacterized protein of Penaeus monodon nudivirus. Real-time qPCR analysis showed that LvWRP was ubiquitously expressed in shrimp tissues, with transcript levels induced in hemocytes upon immune challenge with Vibrio parahaemolyticus, Streptoccocus iniae, lipopolysaccharide (LPS), and WSSV. In addition, RNA interference-mediated knockdown of LvWRP followed by WSSV challenge revealed significant decrease in the transcript levels of WSSV IE1 and VP28 genes coupled with a reduction in WSSV copies in shrimp hemocytes. Moreover, depletion of LvWRP followed by WSSV challenge significantly increased the transcript levels of Vago4 and Vago5 as well as increased the phosphorylation of STAT, while hemocytes apoptosis in terms of caspase 3/7 activity was decreased. These results suggest that LvWRP is important for WSSV replication in shrimp, and therefore one of the vital host factors in WSSV infection.
Gao G
，Lin R
，Tao M
，Aweya JJ
，Yao D
，Ma H
，Li S
，Zhang Y
，Wang F
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The T cell factor, pangolin, from Litopenaeus vannamei play a positive role in the immune responses against white spot syndrome virus infection.
As a downstream interactor of β-catenin, Pangolin which is the homologous protein of the T cell factor/lymphoid enhancer factor (TCF/LEF) in vertebrates is less understood in the research field of immunity. In this study, two isoforms of Litopenaeus vannamei Pangolin (LvPangolin1 and LvPangolin2) were identified. Phylogenetic tree analysis revealed that all of the Pangolin proteins from invertebrates were represented the same lineage. The mRNA expression profiles of the LvPangolin1 and LvPangolin2 genes differed across different tissues. The expression of LvPangolin1 and the amount of LvPangolin1and LvPangolin2 combined (LvPangolinComb) were significantly increased in the haemocyte, intestine and gill but reduced in the hepatopancreas after white spot syndrome virus (WSSV) challenge. The inhibition of LvPangolin1 but not LvPangolinComb significantly reduced the survival rates of L. vannamei after WSSV infection, while significantly higher WSSV viral loads in both LvPangolin1-inhibited and LvPangolinComb-inhibited L. vannamei were observed. Knockdown of LvPangolin by RNAi could distinctly decrease the expression of antimicrobial peptide (AMP) genes and their related transcription factors. All of these results indicate that LvPangolin plays a positive role in the response to WSSV infection and that this may be mediated through regulating the immune signalling pathways which control the expression of AMPs with antiviral abilities.
Zhu L
，Zhang S
，Hou C
，Liang X
，Saif Dehwah MA
，Tan B
，Shi L
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The promotion of cytoskeleton integration and redox in the haemocyte of shrimp Litopenaeus vannamei after the successive stimulation of recombinant VP28.
VP28 protein has been reported to work as a "vaccine" to protect the host from white spot syndrome, but the detailed mechanism of vaccination with VP28 protein in shrimp is still far from well understood. In the present study, whole transcriptomes of shrimp haemocytes were sequenced using the SOLiD4 platform after the successive VP28 stimulation. Eight single-end fragment libraries were constructed and sequenced in the four groups including the VP28-VP28, PBS-VP28, PBS-PBS and BLANK group, and there were 243,949,667 single-end reads with length of 50bp obtained totally, with 14,800 genes further identified. After reads mapping and transcript assembling, 1027, 1539, 1158, 1091 and 1300 genes in five differentially expressed gene lists were obtained in the comparison of VP28-VP28 versus PBS-VP28, VP28-VP28 versus PBS-PBS, VP28-VP28 versus BLANK, PBS-VP28 versus PBS-PBS and PBS-VP28 versus BLANK, respectively. There were 555 differentially expressed genes responsive to the single VP28 stimulation after grouping the PBS-VP28_BLANK and PBS-VP28_PBS-PBS gene lists, and 269 ones responsive to the successive VP28 stimulation after grouping the VP28-VP28_BLANK, VP28-VP28_PBS-PBS and VP28-VP28_PBS-VP28 gene lists. In the GO enrichment analysis of the genes responsive to the single VP28 stimulation, five immune-related GO terms were observed among 14 increased terms, which included defense response to bacterium, response to stimulus, disruption of cells of other organism, killing of cells of other organism and response to bacterium. It was worth noting that the GO terms, response to stimulus and response to stress, were the most common annotation ones which accounted 28.7% and 18.8% of the total differently expressed genes, respectively. For the genes responsive to the successive VP28 stimulation, terms including actin filament-based movement and myosin heavy chain binding were mostly enriched in the Biological Process and Molecular Function category, respectively. In the Cellular Component category, the enriched GO terms were myosin VII complex, myosin V complex, myosin VI complex and myosin II complex. Furthermore, the most abundant GO term was oxidation-reduction process, followed by single-organism transport, neurogenesis and translation for 214 genes only responsive to successive VP28 stimulation. These results collectively indicated that the successive VP28 stimulation could modulate cytoskeleton integration and redox to promote the phagocytosis activity of shrimp haemocytes, which might protect effectively for shrimp against WSSV infection.
Wang L
，Sun X
，Zhou Z
，Zhang T
，Yi Q
，Liu R
，Wang M
，Song L
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