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Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei

Wenfeng Li (UGent) , Lowiese Desmarets (UGent) , Gaëtan De Gryse (UGent) , Sebastiaan Theuns (UGent) , Vo Van Tuan (UGent) , Thuong Van Khuong (UGent) , Peter Bossier (UGent) and Hans Nauwynck (UGent)
(2015) JOURNAL OF GENERAL VIROLOGY. 96(9). p.2844-2854
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Abstract
The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this nnonolayer could be maintained for 10 days with a viability of 90 %. Binding of WSSV to cells reached a maximum (73 +/- 3 % of cells and 4.84 +/- 0.2 virus particles per virus-binding cell) at 120 min at 4 degrees C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosonnes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 degrees C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664- and VP28-positive in their nuclei peaked (50 +/- 4 %) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.
Keywords
SHRIMP, WSSV, INFECTION, VP28

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Chicago
Li, Wenfeng, Lowiese Desmarets, Gaëtan De Gryse, Sebastiaan Theuns, Vo Van Tuan, Thuong Van Khuong, Peter Bossier, and Hans Nauwynck. 2015. “Virus Replication Cycle of White Spot Syndrome Virus in Secondary Cell Cultures from the Lymphoid Organ of Litopenaeus Vannamei.” Journal of General Virology 96 (9): 2844–2854.
APA
Li, Wenfeng, Desmarets, L., De Gryse, G., Theuns, S., Tuan, V. V., Khuong, T. V., Bossier, P., et al. (2015). Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. JOURNAL OF GENERAL VIROLOGY, 96(9), 2844–2854.
Vancouver
1.
Li W, Desmarets L, De Gryse G, Theuns S, Tuan VV, Khuong TV, et al. Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. JOURNAL OF GENERAL VIROLOGY. 2015;96(9):2844–54.
MLA
Li, Wenfeng, Lowiese Desmarets, Gaëtan De Gryse, et al. “Virus Replication Cycle of White Spot Syndrome Virus in Secondary Cell Cultures from the Lymphoid Organ of Litopenaeus Vannamei.” JOURNAL OF GENERAL VIROLOGY 96.9 (2015): 2844–2854. Print.
@article{6872858,
  abstract     = {The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this nnonolayer could be maintained for 10 days with a viability of 90 \%. Binding of WSSV to cells reached a maximum (73 +/- 3 \% of cells and 4.84 +/- 0.2 virus particles per virus-binding cell) at 120 min at 4 degrees C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosonnes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 degrees C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664- and VP28-positive in their nuclei peaked (50 +/- 4 \%) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.},
  author       = {Li, Wenfeng and Desmarets, Lowiese and De Gryse, Ga{\"e}tan and Theuns, Sebastiaan and Tuan, Vo Van and Khuong, Thuong Van and Bossier, Peter and Nauwynck, Hans},
  issn         = {0022-1317},
  journal      = {JOURNAL OF GENERAL VIROLOGY},
  keyword      = {SHRIMP,WSSV,INFECTION,VP28},
  language     = {eng},
  number       = {9},
  pages        = {2844--2854},
  title        = {Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei},
  url          = {http://dx.doi.org/10.1099/vir.0.000217},
  volume       = {96},
  year         = {2015},
}

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