scholarly journals Antilipopolysaccharide Factor Interferes with White Spot Syndrome Virus Replication In Vitro and In Vivo in the Crayfish Pacifastacus leniusculus

2006 ◽  
Vol 80 (21) ◽  
pp. 10365-10371 ◽  
Author(s):  
Haipeng Liu ◽  
Pikul Jiravanichpaisal ◽  
Irene Söderhäll ◽  
Lage Cerenius ◽  
Kenneth Söderhäll
Keyword(s):  
Cell Culture ◽  
White Spot Syndrome Virus ◽  
Pacifastacus Leniusculus ◽  
White Spot ◽  
Freshwater Crayfish ◽  
Hematopoietic Tissue ◽  
Viral Loads ◽  
White Spot Syndrome

ABSTRACT In a study of genes expressed differentially in the freshwater crayfish Pacifastacus leniusculus infected experimentally with the white spot syndrome virus (WSSV), one protein, known as antilipopolysaccharide factor (ALF), was chosen, among those whose transcript levels increased upon viral infection, for further studies. ALF RNA interference (RNAi) experiments in whole animals and in cell cultures indicated that ALF can protect against WSSV infection, since knockdown of ALF by RNAi specifically resulted in higher rates of viral propagation. In a cell culture of hematopoietic tissue (Hpt) from P. leniusculus, quantitative PCR showed that knockdown of ALF by RNAi resulted into WSSV levels that were about 10-fold higher than those treated with control double-stranded RNA (dsRNA). In addition, RNAi experiments with other crayfish genes that had been found to be up-regulated by a WSSV infection did not result in any changes of viral loads. Thus, the cell culture does not respond to dsRNA in a similar manner, as shown earlier for dsRNA injected into shrimp, which gave a higher degree of resistance to WSSV infection. If ALF transcription in whole animals was stimulated by the administration of UV-treated WSSV, a partial protection against a subsequent challenge with the active virus was conferred to the host. This is the first crustacean gene product identified with the capacity to interfere with replication of this important pathogen.

scholarly journals A gC1qR Prevents White Spot Syndrome Virus Replication in the Freshwater Crayfish Pacifastacus leniusculus

2010 ◽  
Vol 84 (20) ◽  
pp. 10844-10851 ◽  
Author(s):  
Apiruck Watthanasurorot ◽  
Pikul Jiravanichpaisal ◽  
Irene Söderhäll ◽  
Kenneth Söderhäll
Keyword(s):  
Cell Culture ◽  
Viral Replication ◽  
White Spot Syndrome Virus ◽  
Pacifastacus Leniusculus ◽  
White Spot ◽  
Tissue Cell ◽  
Freshwater Crayfish ◽  
White Spot Syndrome

ABSTRACT The gC1qR/p32 protein is a multiple receptor for several proteins and pathogens. We cloned a gC1qR homologue in a crustacean, Pacifastacus leniusculus, and analyzed the expression of P. leniusculus C1qR (PlgC1qR) in various tissues. The gC1qR/p32 transcript was significantly enhanced by white spot syndrome virus (WSSV) infection 6 h after viral infection both in vitro in a hematopoietic tissue cell culture (Hpt) and in vivo compared to appropriate controls. Moreover, PlgC1qR silencing in both the Hpt cell culture and live crayfish enhanced the WSSV replication. In addition, by making a recombinant PlgC1qR protein we could show that if this recombinant protein was injected in a crayfish, Pacifastacus leniusculus, followed by injection of WSSV, this significantly reduced viral replication in vivo. Furthermore, if the recombinant PlgC1qR was incubated with Hpt cells and then WSSV was added, this also reduced viral replication. These experiments clearly demonstrate that recombinant PlgC1qR reduce WSSV replication both in vivo and in vitro. The results from a far-Western overlay and glutathione S-transferase pull-down assays showed that PlgC1qR could bind to VP15, VP26, and VP28. Altogether, these results demonstrate a role for PlgC1qR in antiviral activity against WSSV.

scholarly journals White Spot Syndrome Virus Open Reading Frame 222 Encodes a Viral E3 Ligase and Mediates Degradation of a Host Tumor Suppressor via Ubiquitination

2006 ◽  
Vol 80 (8) ◽  
pp. 3884-3892 ◽  
Author(s):  
Fang He ◽  
Beau J. Fenner ◽  
Andrew K. Godwin ◽  
Jimmy Kwang
Keyword(s):  
Tumor Suppressor ◽  
Cell Line ◽  
White Spot Syndrome Virus ◽  
Ovarian Cancer Cell Line ◽  
E3 Ligase ◽  
Cancer Cell Line ◽  
White Spot ◽  
White Spot Syndrome

ABSTRACT We have characterized a white spot syndrome virus (WSSV) RING-H2-type protein, WSSV222, which is involved in ubiquitination. WSSV222 exhibits RING-H2-dependent E3 ligase activity in vitro in the presence of the specific conjugating enzyme UbcH6. Mutations in the RING-H2 domain abolished WSSV222-dependent ubiquitination, revealing the importance of this domain in WSSV222 function. Yeast two-hybrid and pull-down analyses revealed that WSSV222 interacts with a shrimp tumor suppressor-like protein (TSL) sharing 60% identity with human OVCA1. To better characterize the interaction of WSSV222 and TSL in vivo, we established a stable TSL-expressing cell line derived from the human ovarian cancer cell line A2780, where we observed a TSL-dependent prolonged G1 phase. Furthermore, we detected WSSV222-mediated ubiquitination and MG132-sensitive degradation of TSL both in shrimp primary cell culture and in the TSL-expressing cell line. Transient expression of TSL in BHK cells leads to apoptosis, which was rescued by WSSV222. Taken together, our data suggest that WSSV222 acts as an antiapoptosis protein by ubiquitin-mediated proteolysis of TSL to ensure successful WSSV replication in shrimp.

scholarly journals Characterization of white spot syndrome virus replication in in vitro-cultured haematopoietic stem cells of freshwater crayfish, Pacifastacus leniusculus

2006 ◽  
Vol 87 (4) ◽  
pp. 847-854 ◽  
Author(s):  
Pikul Jiravanichpaisal ◽  
Kenneth Söderhäll ◽  
Irene Söderhäll
Keyword(s):  
Virus Replication ◽  
White Spot Syndrome Virus ◽  
Pacifastacus Leniusculus ◽  
White Spot ◽  
Post Inoculation ◽  
Freshwater Crayfish ◽  
Rt Pcr ◽  
Wide Range ◽  
White Spot Syndrome

Replication of White spot syndrome virus (WSSV) was investigated in haematopoietic cells (hpt cells) derived from haematopoietic tissue (hpt) of freshwater crayfish, Pacifastacus leniusculus. Temperature and type of inoculum for virus replication were studied. The cell culture remained viable at a wide range of temperatures ranging from 4 to 25 °C. WSSV replicated in cells, as evidenced by in situ hybridization, RT-PCR and by the presence of virions visualized with an electron microscope. Moreover, the results showed that the infectivity of WSSV to hpt cells is dependent on temperature and a supplemented growth factor (cytokine) astakine. WSSV replicated more rapidly at higher temperatures than at lower temperatures. No virus replication was observed at 4 °C. Detectable WSSV-infected cells were present as early as 36 h post-inoculation, demonstrated by in situ hybridization or RT-PCR of VP28 expression at 25 °C. Hpt cells can survive a few weeks at 25 or 16 °C and longer than several months at 4 °C.

scholarly journals Phycoerythrin from Colaconema sp. Has Immunostimulatory Effects on the Whiteleg Shrimp Litopenaeus vannamei and Increases Resistance to Vibrio parahaemolyticus and White Spot Syndrome Virus

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2371
Author(s):  
Po-Tsang Lee ◽  
Jing Huang ◽  
Chin-Yi Huang ◽  
Zi-Xuan Liu ◽  
Han-Yang Yeh ◽  
...  
Keyword(s):  
Litopenaeus Vannamei ◽  
Vibrio Parahaemolyticus ◽  
White Spot Syndrome Virus ◽  
Expression Profiles ◽  
White Spot ◽  
Shrimp Culture ◽  
White Spot Syndrome ◽  
Whiteleg Shrimp

We investigated whether phycoerythrin (PE), a pigment sourced from marine algae, could act as an immunomodulatory agent in whiteleg shrimp (Litopenaeus vannamei). To this end, PE was extracted and purified from a PE-rich macroalgae, Colaconema sp. Our in vitro analysis demonstrated that PE enhanced prophenoloxidase and phagocytosis activity but inhibited the production of reactive oxygen species in hemocytes. Additionally, the PE signal could be detected using an in vivo imaging system after its injection into the ventral sinus of the cephalothorax of whiteleg shrimp. The expression profiles of fourteen immune-related genes were monitored in hemocytes from whiteleg shrimp injected with 0.30 μg of PE per gram of body weight, and crustin, lysozyme, penaiedin 4, and anti-lipopolysaccharide factor showed up-regulated post-stimulation. The induction of immune genes and enhancement of innate immune parameters by PE may explain the higher survival rates for shrimp that received different doses of PE prior to being challenged with Vibrio parahaemolyticus or white spot syndrome virus compared to controls. Combined, these results show that PE from Colaconema sp. can differentially stimulate the immune response of whiteleg shrimp in vitro and in vivo and could potentially be used as an immunomodulator in shrimp culture.

scholarly journals Role of anti-lipopolysaccharide factor from the black tiger shrimp, Penaeus monodon, in protection from white spot syndrome virus infection

2009 ◽  
Vol 90 (6) ◽  
pp. 1491-1498 ◽  
Author(s):  
Sirinit Tharntada ◽  
Sirikwan Ponprateep ◽  
Kunlaya Somboonwiwat ◽  
Haipeng Liu ◽  
Irene Söderhäll ◽  
...  
Keyword(s):  
Cell Cultures ◽  
White Spot Syndrome Virus ◽  
Penaeus Monodon ◽  
White Spot ◽  
Freshwater Crayfish ◽  
Black Tiger Shrimp ◽  
Tiger Shrimp ◽  
White Spot Syndrome

The anti-lipopolysaccharide factor (ALF) from the black tiger shrimp, Penaeus monodon, has been shown previously to exhibit a broad spectrum of activity against various strains of bacteria and fungi. Herein, the recombinant ALFPm3 (rALFPm3) protein was examined for its role in the defence against white spot syndrome virus (WSSV) infection in haematopoietic (Hpt) cell cultures of the freshwater crayfish, Pacifastacus leniusculus, as well as in live P. monodon shrimps. Incubation of Hpt cell cultures with a mixture of WSSV and rALFPm3 resulted in a dose-dependent decrease in VP28 gene expression levels, compared with those incubated with WSSV alone, with an rALFPm3 IC50 value lower than 2.5 μM. However, pre-treatment of Hpt cells with 5 μM rALFPm3 showed no induced protection against subsequent WSSV infection, whereas the synthetic crayfish ALF peptide could protect cells at a higher concentration (10 μM). The in vivo role of ALFPm3 was examined by injection of P. monodon with WSSV pre-treated with rALFPm3 protein. The results clearly showed that rALFPm3 was able to reduce WSSV propagation and prolong the survival of shrimps.

scholarly journals Identification of a Shrimp E3 Ubiquitin Ligase TRIM50-Like Involved in Restricting White Spot Syndrome Virus Proliferation by Its Mediated Autophagy and Ubiquitination

2021 ◽  
Vol 12 ◽  
Author(s):  
Chao Zhao ◽  
Chao Peng ◽  
Pengfei Wang ◽  
Lulu Yan ◽  
Sigang Fan ◽  
...  
Keyword(s):  
White Spot Syndrome Virus ◽  
Penaeus Monodon ◽  
White Spot ◽  
Positive Role ◽  
Protein Levels ◽  
Mrna And Protein Expression ◽  
White Spot Syndrome ◽  
Critical Components ◽  
Trim Proteins

Most tripartite motif (TRIM) family proteins are critical components of the autophagy machinery and play important roles in host defense against viral pathogens in mammals. However, the roles of TRIM proteins in autophagy and viral infection have not been studied in lower invertebrates, especially crustaceans. In this study, we first identified a TRIM50-like gene from Penaeus monodon (designated PmTRIM50-like), which, after a white spot syndrome virus (WSSV) challenge, was significantly upregulated at the mRNA and protein levels in the intestine and hemocytes. Knockdown of PmTRIM50-like led to an increase in the WSSV quantity in shrimp, while its overexpression led to a decrease compared with the controls. Autophagy can be induced by WSSV or rapamycin challenge and has been shown to play a positive role in restricting WSSV replication in P. monodon. The mRNA and protein expression levels of PmTRIM50-like significantly increased with the enhancement of rapamycin-induced autophagy. The autophagy activity induced by WSSV or rapamycin challenge could be inhibited by silencing PmTRIM50-like in shrimp. Further studies showed that rapamycin failed to induce autophagy or inhibit WSSV replication after knockdown of PmTRIM50-like. Moreover, pull-down and in vitro ubiquitination assays demonstrated that PmTRIM50-like could interact with WSSV envelope proteins and target them for ubiquitination in vitro. Collectively, this study demonstrated that PmTRIM50-like is required for autophagy and is involved in restricting the proliferation of WSSV through its ubiquitination. This is the first study to report the role of a TRIM family protein in virus infection and host autophagy in crustaceans.

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