Environmental concentrations of sulfamethoxazole increase crayfish Pacifastacus leniusculus susceptibility to White Spot Syndrome Virus

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.

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Experimental infection of white spot syndrome virus in freshwater crayfish Pacifastacus leniusculus

Diseases of Aquatic Organisms ◽

10.3354/dao047151 ◽

2001 ◽

Vol 47 ◽

pp. 151-157 ◽

Cited By ~ 78

Author(s):

P Jiravanichpaisal ◽

E Bangyeekhun ◽

K Söderhäll ◽

I Söderhäll

Keyword(s):

Experimental Infection ◽

White Spot Syndrome Virus ◽

Pacifastacus Leniusculus ◽

White Spot ◽

Freshwater Crayfish ◽

White Spot Syndrome

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

Journal of General Virology ◽

10.1099/vir.0.81758-0 ◽

2006 ◽

Vol 87 (4) ◽

pp. 847-854 ◽

Cited By ~ 59

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.

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

Journal of Virology ◽

10.1128/jvi.01101-06 ◽

2006 ◽

Vol 80 (21) ◽

pp. 10365-10371 ◽

Cited By ~ 175

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.

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