title: Ex vivo-generated CD36+ erythroid progenitors are highly permissive to human parvovirus B19 replication.
abstract: The pathogenic parvovirus B19 (B19V) has an extreme tropism for human erythroid progenitor cells. In vitro, only a few erythroid leukemic cell lines (JK-1 and KU812Ep6) or megakaryoblastoid cell lines (UT7/Epo and UT7/Epo-S1) with erythroid characteristics support B19V replication, but these cells are only semipermissive. By using recent advances in generating large numbers of human erythroid progenitor cells (EPCs) ex vivo from hematopoietic stem cells (HSCs), we produced a pure population of CD36(+) EPCs expanded and differentiated from CD34(+) HSCs and assessed the CD36(+) EPCs for their permissiveness to B19V infection. Over more than 3 weeks, cells grown in serum-free medium expanded more than 800,000-fold, and 87 to 96% of the CD36(+) EPCs were positive for globoside, the cellular receptor for B19V. Immunofluorescence (IF) staining showed that about 77% of the CD36(+) EPCs were positive for B19V infection, while about 9% of UT7/Epo-S1 cells were B19V positive. Viral DNA detected by real-time PCR increased by more than 3 logs in CD36(+) EPCs; the increase was 1 log in UT7/Epo-S1 cells. Due to the extensive permissivity of CD36(+) EPCs, we significantly improved the sensitivity of detection of infectious B19V by real-time reverse transcription-PCR and IF staining 100- and 1,000-fold, respectively, which is greater than the sensitivity of UT7/Epo-S1 cell-based methods. This is the first description of an ex vivo method to produce large numbers of EPCs that are highly permissive to B19V infection and replication, offering a cellular system that mimics in vivo infection with this pathogenic human virus.
authors.affiliation: Hematology Branch, NHLBI, NIH, Bldg 10-CRC, Rm 3E-5216, 10 Center Drive, Bethesda, MD 20892, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Wong
authors.forename: Ning
authors.initials: N
authors.lastname: Zhi
authors.forename: Claudia
authors.initials: C
authors.lastname: Filippone
authors.forename: Keyvan
authors.initials: K
authors.lastname: Keyvanfar
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kajigaya
authors.forename: Kevin E
authors.initials: KE
authors.lastname: Brown
authors.forename: Neal S
authors.initials: NS
authors.lastname: Young
mesh_terms: D001483:Base Sequence; D018955:CD36 Antigens / Q000276:immunology*; D002478:Cells, Cultured; D017931:DNA Primers; D015672:Erythroid Precursor Cells / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D016732:Parvovirus B19, Human / Q000502:physiology*; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication
chemical_list: D018955:CD36 Antigens; D017931:DNA Primers