DH82 cells: a macrophage cell line for the replication and study of equineinfectious anemia virus

Authors
Citation
R. Hines et W. Maury, DH82 cells: a macrophage cell line for the replication and study of equineinfectious anemia virus, J VIROL MET, 95(1-2), 2001, pp. 47-56
Citations number
36
Categorie Soggetti
Microbiology
Journal title
JOURNAL OF VIROLOGICAL METHODS
ISSN journal
01660934 → ACNP
Volume
95
Issue
1-2
Year of publication
2001
Pages
47 - 56
Database
ISI
SICI code
0166-0934(200106)95:1-2<47:DCAMCL>2.0.ZU;2-S
Abstract
In vivo, tissue macrophages have been implicated as an important cell for t he replication of equine infectious anemia virus (EIAV). Laboratory investi gations of EIAV;macrophage interactions, however, have been hampered by the laborious blood monocyte isolation procedures. Tn addition. adherent equin e macrophage cultures generally have poor long-term viability and are resis tant to transfection. This report describes an adherent canine macrophage-l ike cell line, DH82. that supports the replication of EIAV. This cell line was easily transfectable and supported EIAV Tat transactivation of the LTR. Electrophoretic mobility shift assays were carried out to determine which transcription factor binding sites within the LTR enhancer region were boun d by DH82 nuclear extracts. IL was found that five: different motifs were o ccupied. The ets motifs that are bound by PU.1 in primary macrophage nuclea r extracts: specifically interacted with DH82 nuclear extracts. In addition , the PEA-2, Lvb and Oct motifs that are occupied by fibroblast nuclear ext racts were also bound by DH82 nuclear extracts. Finally, the methylation-de pendent binding protein (MDBP) site that is: bound by all nuclear extracts investigated to date demonstrated specific interactions with DH82 nuclear e xtracts. The observation that both macrophage-specific and fibroblast-speci fic motifs were utilized by DH82 nuclear extracts suggested that both macro phage-adapted and fibroblast-adapted EIAV could replicate in DH82 cells. In deed, infectivity studies demonstrated that strains of virus that exclusive ly replicate in macrophages can replicate in DH82 cells and fibroblast-adap ted strains of virus can also replicate in these cells. Finally, these cell s could be transfected readily with the EIAV molecular clone, pSPeiav19-2. and virus spread was detected within the culture. In conclusion, this study has identified a useful cell line that should facilitate the study of EIAV expression and replication. (C) 2001 Elsevier Science B.V. All rights rese rved.