1887

Abstract

The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of . The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this monolayer 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 °C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosomes 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 °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. 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.

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2015-09-01
2021-10-20
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References

  1. Assavalapsakul W., Smith D.R., Panyim S. 2003; Propagation of infectious yellow head virus particles prior to cytopathic effect in primary lymphoid cell cultures of Penaeus monodon . Dis Aquat Organ 55:253–258 [View Article][PubMed]
    [Google Scholar]
  2. Chaivisuthangkura P., Tangkhabuanbutra J., Longyant S., Sithigorngul W., Rukpratanporn S., Menasveta P., Sithigorngul P. 2004; Monoclonal antibodies against a truncated viral envelope protein (VP28) can detect white spot syndrome virus (WSSV) infections in shrimp. Sci Asia 30:359–363 [View Article]
    [Google Scholar]
  3. Chavrier P., Parton R.G., Hauri H.P., Simons K., Zerial M. 1990; Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments. Cell 62:317–329 [View Article][PubMed]
    [Google Scholar]
  4. Dantas-Lima J.J., Corteel M., Cornelissen M., Bossier P., Sorgeloos P., Nauwynck H.J. 2013; Purification of white spot syndrome virus by iodixanol density gradient centrifugation. J Fish Dis 36:841–851[PubMed]
    [Google Scholar]
  5. Escobedo-Bonilla C.M., Wille M., Sanz V.A., Sorgeloos P., Pensaert M.B., Nauwynck H.J. 2005; In vivo titration of white spot syndrome virus (WSSV) in specific pathogen-free Litopenaeus vannamei by intramuscular and oral routes. Dis Aquat Organ 66:163–170 [View Article][PubMed]
    [Google Scholar]
  6. Feng Y., Press B., Wandinger-Ness A. 1995; Rab 7: an important regulator of late endocytic membrane traffic. J Cell Biol 131:1435–1452 [View Article][PubMed]
    [Google Scholar]
  7. Flegel T.W. 1997; Special topic review: major viral diseases of the black tiger prawn (Penaeus monodon) in Thailand. World J Microbiol Biotechnol 13:433–442 [View Article]
    [Google Scholar]
  8. Han Q., Li P., Lu X., Guo Z., Guo H. 2013; Improved primary cell culture and subculture of lymphoid organs of the greasyback shrimp Metapenaeus ensis . Aquaculture 410–411:101–113 [View Article]
    [Google Scholar]
  9. Huang Z.J., Kang S.T., Leu J.H., Chen L.L. 2013; Endocytic pathway is indicated for white spot syndrome virus (WSSV) entry in shrimp. Fish Shellfish Immunol 35:707–715 [View Article][PubMed]
    [Google Scholar]
  10. Jiravanichpaisal P., Söderhäll K., Söderhäll I. 2006; Characterization of white spot syndrome virus replication in in vitro-cultured haematopoietic stem cells of freshwater crayfish, Pacifastacus leniusculus . J Gen Virol 87:847–854 [View Article][PubMed]
    [Google Scholar]
  11. Kasornchandra J., Boonyaratpalinn S., Khongpradit R., Akpanithanpong U. 1995; A bacilliform virus, the causative agent of red disease with white patch in black tiger shrimp (Penaeus monodon). Technical Paper 3, National Institute of Coastal Aquaculture, Kaoseng, Songkla, Thailand
    [Google Scholar]
  12. Kasornchandra J., Khongpradit R., Ekpanithanpong U., Boonyaratpalin S. 1999; Progress in the development of shrimp cell cultures in Thailand. Methods Cell Sci 21:231–235 [View Article][PubMed]
    [Google Scholar]
  13. Leu J.H., Tsai J.M., Wang H.C., Wang A.H.J., Wang C.H., Kou G.H., Lo C.F. 2005; The unique stacked rings in the nucleocapsid of the white spot syndrome virus virion are formed by the major structural protein VP664, the largest viral structural protein ever found. J Virol 79:140–149 [View Article][PubMed]
    [Google Scholar]
  14. Leu J.H., Yang F., Zhang X., Xu X., Kou G.H., Lo C.F. 2009; Whispovirus. In Lesser Known Large dsDNA Viruses pp. 197–227 Edited by van Etten J. L. Berlin: Springer; [CrossRef]
    [Google Scholar]
  15. Li F., Li M., Ke W., Ji Y., Bian X., Yan X. 2009; Identification of the immediate-early genes of white spot syndrome virus. Virology 385:267–274 [View Article][PubMed]
    [Google Scholar]
  16. Li W., Nguyen V.T., Corteel M., Dantas-Lima J.J., Thuong K.V., Tuan V.V., Bossier P., Sorgeloos P., Nauwynck H. 2014; Characterization of a primary cell culture from lymphoid organ of Litopenaeus vannamei and use for studies on WSSV replication. Aquaculture 433:157–163 [View Article]
    [Google Scholar]
  17. Li W., Tuan V.V., Thuong K.V., Bossier P., Nauwynck H. 10.1007/s11626-015-9882-2 [Epub ahead of print] Eye extract improves cell migration out of lymphoid organ explants of L. vannamei and viability of the primary cell cultures. In Vitro Cell Dev Biol-Animal [View Article]
    [Google Scholar]
  18. Lin S.T., Chang Y.S., Wang H.C., Tzeng H.F., Chang Z.F., Lin J.Y., Wang C.H., Lo C.F., Kou G.H. 2002; Ribonucleotide reductase of shrimp white spot syndrome virus (WSSV): expression and enzymatic activity in a baculovirus/insect cell system and WSSV-infected shrimp. Virology 304:282–290 [View Article][PubMed]
    [Google Scholar]
  19. Lu C.P., Zhu S., Guo F.S., Wu S.Y. 1997; Electron microscopic observation on a non-occluded baculo-like virus in shrimps. Arch Virol 142:2073–2078 [View Article][PubMed]
    [Google Scholar]
  20. Reed L.J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497
    [Google Scholar]
  21. Sánchez-Paz A. 2010; White spot syndrome virus: an overview on an emergent concern. Vet Res 41:43 [View Article][PubMed]
    [Google Scholar]
  22. Sritunyalucksana K., Wannapapho W., Lo C.F., Flegel T.W. 2006; PmRab7 is a VP28-binding protein involved in white spot syndrome virus infection in shrimp. J Virol 80:10734–10742 [View Article][PubMed]
    [Google Scholar]
  23. Tapay L.M., Lu Y., Brock J.A., Nadala E.C.B. Jr, Loh P.C. 1995; Transformation of primary cultures of shrimp (Penaeus stylirostris) lymphoid (Oka) organ with Simian virus-40 (T) antigen. Proc Soc Exp Biol Med 209:73–78 [View Article][PubMed]
    [Google Scholar]
  24. van Hulten M.C.W., Witteveldt J., Snippe M., Vlak J.M. 2001; White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. Virology 285:228–233 [View Article][PubMed]
    [Google Scholar]
  25. Wang C.H., Yang H.N., Tang C.Y., Lu C.H., Kou G.H., Lo C.F. 2000; Ultrastructure of white spot syndrome virus development in primary lymphoid organ cell cultures. Dis Aquat Organ 41:91–104 [View Article][PubMed]
    [Google Scholar]
  26. Wongteerasupaya C., Vickers J.E., Sriurairatana S., Nash G.L., Akarajamorn A., Boonsaeng V., Panyim S., Tassanakajon A., Withyachumnarnkul B., Flegel T.W. 1995; A non-occcluded systemic baculovirus that occurs in cells of ectodermal and mesodermal origin and causes high mortality in the black tiger prawn Penaeus monodon . Dis Aquat Organ 21:69–77 [View Article]
    [Google Scholar]
  27. Yi G., Wang Z., Qi Y., Yao L., Qian J., Hu L. 2004; Vp28 of shrimp white spot syndrome virus is involved in the attachment and penetration into shrimp cells. J Biochem Mol Biol 37:726–734 [View Article][PubMed]
    [Google Scholar]
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