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First continuous propagation of B19 parvovirus in a cell line
S Shimomura, N Komatsu, N Frickhofen, S Anderson, S Kajigaya and NS Young
Cell Biology Section, National Heart, Lung, and Blood Institute, Bethesda,
MD 20892.
The pathogenic human parvovirus B19 has extreme tropism for human erythroid
progenitor cells and has resisted cultivation in conventional cell lines.
We report first propagation of this virus in an erythropoietin-dependent
strain of a megakaryoblastic leukemia cell line called UT-7. Virus protein
was present in about 5% of cells after 1 week of culture. Appropriate
ratios of major and minor capsid proteins were determined by immunoblot,
and newly synthesized capsid protein was detected by immunoprecipitation of
radioactively labeled cell lysates. High molecular weight monomer and dimer
intermediates were detected by Southern analysis, indicating active viral
replication. Approximately 1,000 genome copies were present per infected
cell, and at the optimal multiplicity of infection 20- to 50- fold more
virus was produced than inoculated. Virus propagation only occurred in UT-7
cells that were adapted to growth in erythropoietin; virus signal was not
detected in UT-7 cells adapted for growth in granulocyte-macrophage
colony-stimulating factor or interleukin-3, even with exposure to
erythropoietin for several days. Infectious virus was detected in cultures
as long as 3 months after inoculation. Despite persistence, there was no
evidence of viral integration on Southern analysis. This cell line may
prove useful for the production of infectious virus and in the analysis of
B19 parvovirus persistence, cytotoxicity, and permissivity.
Volume 79,
Issue 1,
pp. 18-24,
01/01/1992
Copyright © 1992 by The American Society of Hematology
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