Characterization of a new fibroblast cell line from a tail fin of red sea bream, Pagrus major, and phylogenetic relationships of a recent RSIV isolate in Japan

Red sea bream iridovirus (RSIV) is a causative agent of red sea brearn iridoviral disease (RSIVD) in marine fish species in Japan. Fibroblast cells were developed from a tail fin of red sea bream, Pagrus major, and then underwent single cell cloning. The successful cloned cells were named CRF-1 cells. Most CRF-1 cells had a normal diploid karyotype with 2n = 48 by chromosomal analysis. RSIV-infected CRF-1 cells showed typical morphological changes that were associated with apoptosis by EGFP-annexin V staining. The serial viral passages were successful in CRF-1 cells but failed in BF-2 cells as judged by MTT assay. The expression of three genes obviously decreased in BF-2 cells compared with CRF-1 cells and finally was below detectable level. Because the expression of 59 1 R gene showed the fastest decrease among three transcripts, the Suppression of IE transcript may be responsible for the restricted replication in BF-2 cells. MCP and ATPase phylogenetic trees showed that RSIV strain U-1 belongs to a distinct group from RSIV strain ehime-1. Therefore, possibly recent epizootics of RSIVD in Japan do not originate directly from RSIV strain ehime-1. Taken together, this study confirmed that RSIV strain U-1 is more closely related to Korean RSIV isolates. (C) 2007 Elsevier B.V. All rights reserved.