Enhanced Cell-Based Detection of Parvovirus B19V Infectious Units According to Cell Cycle Status

Abstract

Human parvovirus B19 (B19V) causes various human diseases, ranging from childhood benign infection to arthropathies, severe anemia and fetal hydrops, depending on the health state and hematological status of the patient. To counteract B19V blood-borne contamination, evaluation of B19 DNA in plasma pools and viral inactivation/removal steps are performed, but nucleic acid testing does not correctly reflect B19V infectivity. There is currently no appropriate cellular model for detection of infectious units of B19V. We describe here an improved cell-based method for detecting B19V infectious units by evaluating its host transcription. We evaluated the ability of various cell lines to support B19V infection. Of all tested, UT7/Epo cell line, UT7/Epo-STI, showed the greatest sensitivity to B19 infection combined with ease of performance. We generated stable clones by limiting dilution on the UT7/Epo-STI cell line with graduated permissiveness for B19V and demonstrated a direct correlation between infectivity and S/G2/M cell cycle stage. Two of the clones tested, B12 and E2, reached sensitivity levels higher than those of UT7/Epo-S1 and CD36+ erythroid progenitor cells. These findings highlight the importance of cell cycle status for sensitivity to B19V, and we propose a promising new straightforward cell-based method for quantifying B19V infectious units.

A Functional Minigenome of Parvovirus B19

“…However, generation of transducing virus was not shown in our experiments, thus constituting the major limit of the present work. All transfection experiments have been conducted in the UT7/EpoS1 cell line, which is appropriate for research on B19V [36], but at the expense of a very low transfection efficiency. When transfected with a complete genomic insert, de novo produced virus can be subsequently amplified in primary EPCs to yield infectious virus at high titre.…”