Adeno-associated virus 2-mediated transduction and erythroid lineage- restricted expression from parvovirus B19p6 promoter in primary human hematopoietic progenitor cells

Chandrika Kurpad, Pinku Mukherjee, X. U.Shan Wang, Selvarangan Ponnazhagan, Linglin Li, Mervin C. Yoder, Arun Srivastava

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Abstract

Human parvovirus B19 gene expression from the viral p6 promoter (B19p6) is restricted to primary human hematopoietic cells undergoing erythroid differentiation. We have demonstrated that expression from this promoter does not occur in established human erythoid cell lines in the context of a recombinant parvovirus genome (Ponnazhagan et al. J Virol 69:8096-8101, 1995). However, abundant expression from this promoter can be readily detected in primary human bone marrow cells (Wang et al. Proc Natl Acad Sci USA 92:12416-12420, 1995; Ponnazhagan et al. J Gen Virol 77:1111-1122, 1996). In the present studies, we investigated the pattern of expression from the B19p6 promoter in primary human bone marrow-derived CD34+ HPC undergoing differentiation into myeloid and erythroid lineages. CD34+ cells were transduced with recombinant adeno-associated virus 2 (AAV) vectors containing the β-galactosidase (lacZ) gene under the control of the following promoters/enhancers: the cytomegalovirus promoter (vCMVp-lacZ), B19p6 promoter (vB19p6-lacZ), B19p6 promoter with an upstream erythroid cell- specific enhancer element (HS-2) from the locus control region (LCR) from the human β-globin gene cluster (vHS2-B19p6-lacZ), and the human β-globin gene promoter with the HS-2 enhancer (vHS2-βp-lacZ). Transgene expression was evaluated either 48 h after infection or following erythroid differentiation in vitro for 3 weeks. Whereas high-level expression from the CMV promoter 48 h after infection diminished with time, low-level expression from the B19p6 and the β-globin promoters increased significantly following erythroid differentiation. Furthermore, in HPC assays, there was no significant difference in the level of expression from the CMV promoter in myeloid or erythroid cell-derived colonies. Expression from the B19p6 and the β-globin promoters, on the other hand, was restricted to erythroid cell colonies. These data further corroborate that the B19p6 promoter is erythroid cell- specific and suggest that the recombinant AAV-B19 hybrid vectors may prove useful in gene therapy of human hemoglobinopathies in general and sickle cell anemia and β-thalassemia in particular.

Original languageEnglish (US)
Pages (from-to)585-592
Number of pages8
JournalJournal of Hematotherapy and Stem Cell Research
Volume8
Issue number6
DOIs
StatePublished - Dec 1 1999

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ASJC Scopus subject areas

  • Immunology
  • Hematology

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