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Antilipopolysaccharide factor interferes with white spot syndrome virus replication in vitro and in vivo in the crayfish Pacifastacus leniusculus
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
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2006 (English)In: Journal of Virology, ISSN 0022-538X, E-ISSN 1098-5514, Vol. 80, no 21, 10365-10371 p.Article in journal (Refereed) Published
Abstract [en]

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 AILF 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 LTV-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.

Place, publisher, year, edition, pages
2006. Vol. 80, no 21, 10365-10371 p.
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:uu:diva-97410DOI: 10.1128/JVI.01101-06ISI: 000241606100008PubMedID: 17041217OAI: oai:DiVA.org:uu-97410DiVA: diva2:172351
Available from: 2008-08-18 Created: 2008-08-18 Last updated: 2017-12-14Bibliographically approved
In thesis
1. Functional Studies of Some Immune Relevant Genes in a Crustacean
Open this publication in new window or tab >>Functional Studies of Some Immune Relevant Genes in a Crustacean
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The freshwater crayfish, Pacifastacus leniusculus, mounts a strong innate immune response against microbes such as viruses and bacteria. In this thesis, a novel RNA interference (RNAi) method mediated with histone H2A was developed and applied in crayfish hematopoietic tissue cell cultures for gene functional studies. Further, the interactions between host (crayfish) and pathogens (white spot syndrome virus and Aeromonas hydrophila, respectively) were studied using RNAi technology in live animals. An antilipopolysaccharide factor isolated from viral challenged crayfish by suppression subtractive hybridization was shown to interfere with the propagation of white spot syndrome virus both in vivo and in vitro in crayfish, suggesting an important role of this factor in antiviral defense. Besides, RNAi of phenoloxidase, a critical immune effector involved in melanization, revealed that phenoloxidase activity is necessary for crayfish immune defense against a highly pathogenic bacterial infection in crayfish. In addition, RNAi was also employed to study a marker protein gene involved in hemocyte maturation in crayfish. Taken together, these studies may provide more insights into the immune responses against pathogen invasion as well as hemocyte ontogenesis in crustaceans.

Place, publisher, year, edition, pages
Uppsala: Universitetsbiblioteket, 2008. 61 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 453
Keyword
anti-LPS factor, crayfish, hemocyte, innate immunity, melanization, phenoloxidase, RNA interference, WSSV
National Category
Biological Sciences
Identifiers
urn:nbn:se:uu:diva-9194 (URN)978-91-554-7252-8 (ISBN)
Public defence
2008-09-12, Lindahlsalen, EBC, Norbyvägen 18A, Uppsala, 10:00 (English)
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Available from: 2008-08-18 Created: 2008-08-18 Last updated: 2009-04-05Bibliographically approved

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Jiravanichpaisal, PikulSöderhäll, IreneCerenius, LageSöderhäll, Kenneth

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