Development of two cell lines from Epinephelus coioides brain tissue for characterization of betanodavirus and megalocytivirus infectivity and propagation

作者：Wen C M^*; Lee C W; Wang C S; Cheng Y H; Huang H Y

来源：Aquaculture, 2008, 278(1-4): 14-21.

DOI：10.1016/j.aquaculture.2008.03.020

• astroglia
• piscine cell line
• grouper
• iridovirus
• Mx protein
• nervous necrosis virus

Betanodaviruses and megalocytiviruses are the causative agents of viral nervous necrosis and iridoviral disease, respectively, among marine species farmed in Taiwan. Because there are few cell lines susceptible to these viruses, we wished to identify additional lines with which to study viral pathology and epidemiology. Thus, we established two clonal cell lines designated GBC1 and GBC4 from the brain of an orange-spotted grouper, Epinephelus coioides. Both cell lines grew well in Leibovitz's L-15 medium supplemented with 5% (GBC1) or 10% (GBC4) fetal bovine serum at temperatures between 20 degrees C and 35 degrees C. Cytokeratin immunofluorescence staining revealed that both cell lines were of epithelial origin. GBC4 cells expressed glial fibril acidic protein suggesting, that they are astroglial lineage cells. The modal diploid chromosome number was 44 and 48 for GBC1 and GBC4, respectively. GBC1 cells were highly susceptible to grouper nerve necrosis virus (GNNV) and yielded titers of 10(10) TCID50 ml(-1) but were non-susceptible to the giant seaperch iridovirus (GSIV). By contrast, GBC4 cells were susceptible to GSIV with titers approaching 10(9) TCID50 Ml(-1) whereas GNNV infection only yielded titers of 10(epsilon); TCID50 ml(-1). GNNV propagated in GBC1 across a wide range of temperatures (15-37 degrees C) whereas in GBC4, GSIV propagated over 15-30 degrees C. Induction of the Mx protein upon GNNV infection occurred in GBC4 but not in GBC1, suggesting that the Mx protein inhibits virus production. Our results suggest that these two cell lines provide a valuable tool for the isolation and investigation of betanodavirus and megalocytivirus.

• 出版日期2008-6-10