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Journal of Virology, March 2008, p. 2470-2476, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.02247-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Ex Vivo-Generated CD36+ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication{triangledown}

Susan Wong, Ning Zhi,* Claudia Filippone,{ddagger} Keyvan Keyvanfar, Sachiko Kajigaya, Kevin E. Brown,{dagger},§ and Neal S. Young{dagger}

Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland

Received 16 October 2007/ Accepted 14 December 2007

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.

* Corresponding author. Mailing address: Hematology Branch, NHLBI, NIH, Bldg 10-CRC, Rm 3E-5216, 10 Center Drive, Bethesda, MD 20892. Phone: (301) 451-7137. Fax: (301) 496-8396. E-mail: zhin{at}nhlbi.nih.gov

{triangledown} Published ahead of print on 26 December 2007.

{ddagger} Present address: Department of Clinical and Experimental Medicine, Division of Microbiology, University of Bologna, Bologna, Italy.

{dagger} Neal S. Young and Kevin E. Brown equally supervised the project.

§ Present address: Virus Reference Department, Centre for Infections, Health Protection Agency, 61 Colindale Avenue, London, NW9 5EQ, United Kingdom.

Journal of Virology, March 2008, p. 2470-2476, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.02247-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Guan, W., Wong, S., Zhi, N., Qiu, J. (2009). The Genome of Human Parvovirus B19 Can Replicate in Nonpermissive Cells with the Help of Adenovirus Genes and Produces Infectious Virus. J. Virol. 83: 9541-9553 [Abstract] [Full Text]  
  • Guan, W., Cheng, F., Yoto, Y., Kleiboeker, S., Wong, S., Zhi, N., Pintel, D. J., Qiu, J. (2008). Block to the Production of Full-Length B19 Virus Transcripts by Internal Polyadenylation Is Overcome by Replication of the Viral Genome. J. Virol. 82: 9951-9963 [Abstract] [Full Text]  
  • Pillet, S., Fichelson, S., Morinet, F., Young, N. S., Zhi, N., Wong, S. (2008). Human B19 Erythrovirus In Vitro Replication: What's New?. J. Virol. 82: 8951-8953 [Full Text]  


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