title: Propagation of classical swine fever virus in vitro circumventing heparan sulfate-adaptation.
abstract: Amplification of natural virus isolates in permanent cell lines can result in adaptation, in particular enhanced binding to heparan sulfate (HS)-containing glycosaminoglycans present on most vertebrate cells. This has been reported for several viruses, including the pestivirus classical swine fever virus (CSFV), the causative agent of a highly contagious hemorrhagic disease in pigs. Propagation of CSFV in cell culture is essential in virus diagnostics and research. Adaptation of CSFV to HS-binding has been related to amino acid changes in the viral E(rns) glycoprotein, resulting in viruses with altered replication characteristics in vitro and in vivo. Consequently, a compound blocking the HS-containing structures on cell surfaces was employed to monitor conversion from HS-independency to HS-dependency. It was shown that the porcine PEDSV.15 cell line permitted propagation of CSFV within a limited number of passages without adaptation to HS-binding. The selection of HS-dependent CSFV mutants was also prevented by propagation of the virus in the presence of DSTP 27. The importance of these findings can be seen from the altered ratio of cell-associated to secreted virus upon acquisition of enhanced HS-binding affinity, a phenotype proposed previously to be related to virulence in the natural host.
authors.affiliation: Institute of Virology and Immunoprophylaxis, Sensemattstrasse 293, CH-3147 Mittelhäusern, Switzerland.
authors.forename: Rita
authors.initials: R
authors.lastname: Eymann-Häni
authors.forename: Immanuel
authors.initials: I
authors.lastname: Leifer
authors.forename: Kenneth C
authors.initials: KC
authors.lastname: McCullough
authors.forename: Artur
authors.initials: A
authors.lastname: Summerfield
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ruggli
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D001011:Aorta / Q000166:cytology / Q000821:virology; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D042783:Endothelial Cells / Q000821:virology; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000821:virology; D010069:Oxadiazoles / Q000138:chemical synthesis / Q000494:pharmacology*; D011743:Pyrimidines / Q000138:chemical synthesis / Q000494:pharmacology*; D013552:Swine; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010948:Viral Plaque Assay / Q000379:methods; D014779:Virus Replication / Q000187:drug effects
chemical_list: D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D010069:Oxadiazoles; D011743:Pyrimidines; D014759:Viral Envelope Proteins; C476527:N,N'-bis(1-oxido(1,2,5)oxadiazolo(3,4-d)pyrimidin-7-yl)-3,12-diaza-6,9-diazonia(5,2,5,2)dispirohexadecane dichloride; C087086:glycoprotein E2, classical swine fever virus