White spot syndrome virus (WSSV) interaction with crayfish haemocytes |
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| Authors: | Jiravanichpaisal Pikul Sricharoen Siripavee Söderhäll Irene Söderhäll Kenneth |
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| Institution: | 1. Department of Comparative Physiology, Evolutionary Biology Centre, Uppsala University, Norbyvägen 18A, SE-752 36 Uppsala, Sweden;2. Department of Fishery, Faculty of Agricultural, Khon Kaen University, Khon Kaen 40002, Thailand;1. Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China;2. Center for Proteomics, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 500006, China;1. Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, Qingdao 266003, PR China;2. Laboratory for Marine Fisheries and Aquaculture, Qingdao National Laboratory for Marine Science and Technology, No. 1 Wenhai Road, Aoshanwei Town, Jimo, Qingdao, PR China;1. Department of Aquaculture, Faculty of Fisheries and Marine Sciences, IPB University, Bogor, West Java, 16680, Indonesia;2. National Research and Innovation Agency (BRIN) of the Republic of Indonesia, Jl. M.H Thamrin, Central Jakarta, 10340, Indonesia;1. College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, Henan 450002, China;2. Henan Key Laboratory of Innovation and Utilization of Grassland Resources, Zhengzhou, Henan 450002, China;3. Henan Forage Engineering Technology Research Center, Zhengzhou, Henan 450002, China;4. College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan 450002, China;1. State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China;2. Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Fujian Engineering Laboratory of Marine Bioproducts and Technology, Xiamen 361102, Fujian, PR China;1. Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Salaya, Nakhonpathom 73170, Thailand;2. Shrimp-Virus Interaction Laboratory (ASVI), National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Rama VI Rd., Bangkok, Thailand |
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| Abstract: | WSSV particles were detected in separated granular cells (GCs) and semigranular cells (SGCs) by in situ hybridisation from WSSV-infected crayfish and the prevalence of WSSV-infected GCs was 5%, whereas it was 22% in SGCs. This indicates that SGCs are more susceptible to WSSV and that this virus replicated more rapidly in SGCs than in GCs and as a result the number of SGCs gradually decreased from the blood circulation. The effect of haemocyte lysate supernatant (HLS), containing the degranulation factor (peroxinectin), phorbol 12-myristate 13-acetate (PMA), the Ca(2+) ionophore A23187 on GCs from WSSV-infected and sham-injected crayfish was studied. The results showed that the percentage of degranulated GCs of WSSV-infected crayfish treated with HLS or PMA was significantly lower than that in the control, whereas no significant difference was observed when treated with the Ca(2+) ionophore. It was previously shown that peroxinectin and PMA have a degranulation effect via intracellular signalling involving protein kinase C (PKC), whereas the Ca(2+) ionophore uses an alternative pathway. HLS treatment of GCs and SGCs from WSSV-infected crayfish results in three different morphological types: non-spread, spread and degranulated cells. The non-spread cell group from both GCs and SGCs after treatment with HLS had more WSSV positive cells than degranulated cells, when detected by in situ hybridisation. Taken together, it is reasonable to speculate that the PKC pathway might be affected during WSSV infection. Another interesting phenomenon was that GCs from non-infected crayfish exhibited melanisation, when incubated in L-15 medium, while no melanisation was found in GCs of WSSV-infected crayfish. However, the phenoloxidase activities of both sham- and WSSV-injected crayfish in HLS were the same as well as proPO expression as detected by RT-PCR. This suggests that the WSSV inhibits the proPO system upstream of phenoloxidase or simply consumes the native substrate for the enzyme so that no activity is shown. The percentage of apoptotic haemocytes in WSSV-infected crayfish was very low, but it was significantly higher than that in the sham-injected crayfish on day 3 or 5 post-infection. The TEM observation in haematopoietic cells (hpt cells) suggests that WSSV infect specific cell types in haematopoietic tissue and non-granular hpt cells seem more favourable to WSSV infection. |
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| Keywords: | White spot syndrome virus (WSSV) Haemocyte lysate supernatant (HLS) Ca2+ ionophore A23187 Phorbol 12-myristate 13-acetate (PMA) Crayfish haemocytes Haematopoietic tissue |
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