title: Development of a walleye spleen stromal cell line sensitive to viral hemorrhagic septicemia virus (VHSV IVb) and to protection by synthetic dsRNA.
abstract: A cell line, WE-spleen6, has been developed from the stromal layer of primary spleen cell cultures. On conventional plastic, WE-spleen6 cells had a spindle-shaped morphology at low cell density but grew to become epithelial-like at confluency. On the commercial extracellular matrix (ECM), Matrigel, the cells remained spindle-shaped and formed lumen-like structures. WE-spleen6 cells had intermediate filament protein, vimentin and the ECM protein, collagen I, but not smooth muscle α-actin (SMA) and von Willebrand factor (vWF) and lacked alkaline phosphatase and phagocytic activities. WE-spleen6 was more susceptible to infection with VHSV IVb than a fibroblast and epithelial cell lines from the walleye caudal fin, WE-cfin11f and WE-cfin11e, respectively. Viral transcripts and proteins appeared earlier in WE-spleen6 cultures as did cytopathic effect (CPE) and significant virus production. The synthetic double-stranded RNA (dsRNA), polyinosinic: polycytidylic acid (pIC), induced the antiviral protein Mx in both cell lines. Treating WE-spleen6 cultures with pIC prior to infection with VHSV IVb inhibited the early accumulation of viral transcripts and proteins and delayed the appearance of CPE and significant viral production. Of particular note, pIC caused the disappearance of viral P protein 2 days post infection. WE-spleen6 should be useful for investigating the impact of VHSV IVb on hematopoietic organs and the actions of pIC on the rhabdovirus life cycle.
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
authors.forename: Nguyen T K
authors.initials: NT
authors.lastname: Vo
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
authors.forename: Aaron W
authors.initials: AW
authors.lastname: Bender
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
authors.forename: Dustin A
authors.initials: DA
authors.lastname: Ammendolia
authors.affiliation: Ontario Veterinary College, Pathobiology, University of Guelph, Guelph, ON N2G 2W1, Canada.
authors.forename: John S
authors.initials: JS
authors.lastname: Lumsden
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada.
authors.forename: Brian
authors.initials: B
authors.lastname: Dixon
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada. Electronic address: ncbols@uwaterloo.ca.
authors.forename: Niels C
authors.initials: NC
authors.lastname: Bols
mesh_terms: D000818:Animals; D002460:Cell Line; D005393:Fish Diseases / Q000821:virology*; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D031941:Hemorrhagic Septicemia, Viral / Q000821:virology*; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism; D029141:Novirhabdovirus / Q000502:physiology*; D010471:Perches; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D011070:Poly I-C / Q000494:pharmacology; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism; D013154:Spleen / Q000821:virology*; D017154:Stromal Cells / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D064547:Myxovirus Resistance Proteins; D010750:Phosphoproteins; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D011070:Poly I-C
keywords: Mx; Poly IC; Spleen cell line; VHSV; Walleye