Adenoviral vector-mediated gene transfer to primitive human hematopoietic progenitor cells

Assessment of transduction and toxicity in long-term culture

Karen L. MacKenzie, Neil R. Hackett, Ronald Crystal, Malcolm A S Moore

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Adenoviral gene transfer to primitive hematopoietic progenitor cells (HPCs) would be useful in gene therapy applications where transient, high- level transgene expression is required. In the present investigations, we have used an adenoviral vector expressing the green fluorescent protein (AdGFP) to quantity transduction of primitive HPCs and assess adenoviral- associated toxicity in long-term culture. Here we show that a cytokine cocktail protects mass populations of CD34+ cells and primary colony forming unit-cultures (CFU-Cs) from toxicity, enabling transduction of up to 79% of CD34+ cells. Transduction of CFU-Cs and more primitive HPCs was quantified following fluorescence activated cell sorting for green flourescence protein expression. Our results demonstrate transduction of 45% of primary CFU-Cs, 33% of week-5 cobblestone area forming cells (CAFCs), and 18% of week-5 CFU- Cs. However, AdGFP infection inhibited proliferation of more primitive cells. Although there was no apparent quantitative change in week-5 CAFCs, the clonogenic capacity of week-5 AdGFP-infected cells was reduced by 40% (P < .01) when compared with mock-infected cells. Adenoviral toxicity specifically affected the transduced subset of primitive HPCs. Transduction of primitive cells is therefore probably underestimated by week-5 CFU-Cs and more accurately indicated by week-5 CAFCs. These studies provide the first functional and quantitative evidence of adenoviral transduction of primitive HPCs. However, further investigations will be necessary to overcome adenoviral toxicity toward primitive HPCs before adenoviral vectors can be considered a safe option for gene therapy. (C) 2000 by The American Society of Hematology.

Original languageEnglish
Pages (from-to)100-108
Number of pages9
JournalBlood
Volume96
Issue number1
Publication statusPublished - 1 Jul 2000
Externally publishedYes

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Gene transfer
Hematopoietic Stem Cells
Toxicity
Green Fluorescent Proteins
Gene therapy
Genes
Stem Cells
Sorting
Genetic Therapy
Fluorescence
Cells
Cytokines
Transgenes
Proteins
Flow Cytometry

ASJC Scopus subject areas

  • Hematology

Cite this

Adenoviral vector-mediated gene transfer to primitive human hematopoietic progenitor cells : Assessment of transduction and toxicity in long-term culture. / MacKenzie, Karen L.; Hackett, Neil R.; Crystal, Ronald; Moore, Malcolm A S.

In: Blood, Vol. 96, No. 1, 01.07.2000, p. 100-108.

Research output: Contribution to journalArticle

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