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Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell

Vo Van Tuan (UGent) , Gaëtan De Gryse (UGent) , Thuong Van Khuong (UGent) , Peter Bossier (UGent) and Hans Nauwynck (UGent)
(2016) JOURNAL OF FISH DISEASES. 39(12). p.1477-1493
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Abstract
Little is known about the innate antiviral defence of shrimp haemocytes. In this context, the haemocytes of penaeid shrimp Litopenaeus vannamei (Boone) were separated by iodixanol density gradient centrifugation into five subpopulations (sub): sub 1 (hyalinocytes), sub 2 and 3 (prohyalinocytes), sub 4 (semigranulocytes) and sub 5 (granulocytes) and exposed to beads, white spot syndrome virus (WSSV) and ultraviolet (UV)-killed WSSV. In a first experiment, the uptake of beads, white spot syndrome virus (WSSV) and UV-killed WSSV by these different haemocyte subpopulations was investigated using confocal microscopy. Only haemocytes of sub 1, 4 and 5 were internalizing beads, WSSV and UV-killed WSSV. Beads were engulfed by a much larger percentage of cells (91.2 in sub 1; 84.1 in sub 4 and 58.1 in sub 5) compared to WSSV (9.6 in sub 1; 10.5 in sub 4 and 7.9 in sub 5) and UV-killed WSSV (12.9 in sub 1; 13.3 in sub 4; and 11.8 in sub 5). In a second experiment, it was shown that upon internalization, WSS virions lost their envelope most probably by fusion with the cellular membrane of the endosome (starting between 30 and 60min post-inoculation) and that afterwards the capsid started to become disintegrated (from 360min post-inoculation). Expression of new viral proteins was not observed. Incubation of haemocyte subpopulations with WSSV but not with UV-killed WSSV and polystyrene beads resulted in a significant drop in haemocyte viability. To find the underlying mechanism, a third experiment was performed in which haemocyte subpopulations were exposed to a short WSSV DNA fragment (VP19) and CpG ODNs. These small DNA fragments induced cell death. In conclusion, WSSV is efficiently internalized by hyalinocytes, semigranulocytes and granulocytes, after which the virus loses its envelope; as soon as the capsids start to disintegrate, cell death is activated, which in part may be explained by the exposure of viral DNA to cellular-sensing molecules.
Keywords
apoptosis, Litopenaeus vannamei, phagocytosis, subpopulations, virus, POLYMERASE-CHAIN-REACTION, DENSITY GRADIENT CENTRIFUGATION, BACULOVIRUS WSBV, CRAYFISH HEMOCYTES, IMMUNE-RESPONSES, WSSV INFECTION, MONODON, REPLICATION, ATTACHMENT, SEPARATION

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MLA
Tuan, Vo Van et al. “Kinetic Analysis of Internalization of White Spot Syndrome Virus by Haemocyte Subpopulations of Penaeid Shrimp, Litopenaeus Vannamei (Boone), and the Outcome for Virus and Cell.” JOURNAL OF FISH DISEASES 39.12 (2016): 1477–1493. Print.
APA
Tuan, V. V., De Gryse, G., Khuong, T. V., Bossier, P., & Nauwynck, H. (2016). Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell. JOURNAL OF FISH DISEASES, 39(12), 1477–1493.
Chicago author-date
Tuan, Vo Van, Gaëtan De Gryse, Thuong Van Khuong, Peter Bossier, and Hans Nauwynck. 2016. “Kinetic Analysis of Internalization of White Spot Syndrome Virus by Haemocyte Subpopulations of Penaeid Shrimp, Litopenaeus Vannamei (Boone), and the Outcome for Virus and Cell.” Journal of Fish Diseases 39 (12): 1477–1493.
Chicago author-date (all authors)
Tuan, Vo Van, Gaëtan De Gryse, Thuong Van Khuong, Peter Bossier, and Hans Nauwynck. 2016. “Kinetic Analysis of Internalization of White Spot Syndrome Virus by Haemocyte Subpopulations of Penaeid Shrimp, Litopenaeus Vannamei (Boone), and the Outcome for Virus and Cell.” Journal of Fish Diseases 39 (12): 1477–1493.
Vancouver
1.
Tuan VV, De Gryse G, Khuong TV, Bossier P, Nauwynck H. Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell. JOURNAL OF FISH DISEASES. 2016;39(12):1477–93.
IEEE
[1]
V. V. Tuan, G. De Gryse, T. V. Khuong, P. Bossier, and H. Nauwynck, “Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell,” JOURNAL OF FISH DISEASES, vol. 39, no. 12, pp. 1477–1493, 2016.
@article{8504195,
  abstract     = {Little is known about the innate antiviral defence of shrimp haemocytes. In this context, the haemocytes of penaeid shrimp Litopenaeus vannamei (Boone) were separated by iodixanol density gradient centrifugation into five subpopulations (sub): sub 1 (hyalinocytes), sub 2 and 3 (prohyalinocytes), sub 4 (semigranulocytes) and sub 5 (granulocytes) and exposed to beads, white spot syndrome virus (WSSV) and ultraviolet (UV)-killed WSSV. In a first experiment, the uptake of beads, white spot syndrome virus (WSSV) and UV-killed WSSV by these different haemocyte subpopulations was investigated using confocal microscopy. Only haemocytes of sub 1, 4 and 5 were internalizing beads, WSSV and UV-killed WSSV. Beads were engulfed by a much larger percentage of cells (91.2 in sub 1; 84.1 in sub 4 and 58.1 in sub 5) compared to WSSV (9.6 in sub 1; 10.5 in sub 4 and 7.9 in sub 5) and UV-killed WSSV (12.9 in sub 1; 13.3 in sub 4; and 11.8 in sub 5). In a second experiment, it was shown that upon internalization, WSS virions lost their envelope most probably by fusion with the cellular membrane of the endosome (starting between 30 and 60min post-inoculation) and that afterwards the capsid started to become disintegrated (from 360min post-inoculation). Expression of new viral proteins was not observed. Incubation of haemocyte subpopulations with WSSV but not with UV-killed WSSV and polystyrene beads resulted in a significant drop in haemocyte viability. To find the underlying mechanism, a third experiment was performed in which haemocyte subpopulations were exposed to a short WSSV DNA fragment (VP19) and CpG ODNs. These small DNA fragments induced cell death. In conclusion, WSSV is efficiently internalized by hyalinocytes, semigranulocytes and granulocytes, after which the virus loses its envelope; as soon as the capsids start to disintegrate, cell death is activated, which in part may be explained by the exposure of viral DNA to cellular-sensing molecules.},
  author       = {Tuan, Vo Van and De Gryse, Gaëtan and Khuong, Thuong Van and Bossier, Peter and Nauwynck, Hans},
  issn         = {0140-7775},
  journal      = {JOURNAL OF FISH DISEASES},
  keywords     = {apoptosis,Litopenaeus vannamei,phagocytosis,subpopulations,virus,POLYMERASE-CHAIN-REACTION,DENSITY GRADIENT CENTRIFUGATION,BACULOVIRUS WSBV,CRAYFISH HEMOCYTES,IMMUNE-RESPONSES,WSSV INFECTION,MONODON,REPLICATION,ATTACHMENT,SEPARATION},
  language     = {eng},
  number       = {12},
  pages        = {1477--1493},
  title        = {Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell},
  url          = {http://dx.doi.org/10.1111/jfd.12482},
  volume       = {39},
  year         = {2016},
}

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