Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer

Karen Kearns, Ingrid Bahner, Gerhard Bauer, Sui Fang Wei, Penelope Valdez, Sarah Wheeler, Lin Woods, Robert Miller, Dennis Casciato, Jeffrey Galpin, Joseph Church, Donald B. Kohn

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Bone marrow samples from 21 human immunodeficiency virus type 1 (HIV-1)-infected subjects were evaluated for their suitability for retrovirus-mediated gene transduction with anti-HIV-1 genes. The percentages of CD34+ cells that could be isolated from the mononuclear fraction of bone marrow samples were determined. Fifteen of the 21 marrow samples had normal percentages of CD34+ cells isolated by immunomagnetic methods. All seven donors with CD4 counts > 100/mm3 had normal percentages of CD34+ cells; of 14 patients with low CD4 cell counts (< 100/mm3), 5 had reduced and 9 had normal percentages of CD34+ cells. Samples of the marrow were plated in a methylcellulose colony-forming unit (CFU) assay to determine the clonogenic capacity of the progenitor cells. Overall,the marrow samples from HIV-infected donors showed a 44% reduction in CFU derived from the mononuclear cell fraction and a 75% reduction in CFU derived from the isolated CD34+ cell fraction, when compared to marrow samples from uninfected donors. Isolated CD34+ cells were transduced with retroviral vectors containing various anti-HIV-1 genes to determine their susceptibility to gene transfer. Transduction of the clonogenic CD34+ cells by retroviral vectors did not differ among marrow samples from 13 HIV-1+ donors and 9 uninfected donors. Long-term bone marrow cultures established from the transduced CD34+ cells demonstrated equivalent survival of clonogenic progenitor cells from both HIV-1-infected and uninfected marrows. Toxicity from expression of the anti-HIV-1 genes was not observed; the percentages of clonogenic progenitor cells that survived in cultures transduced by vectors carrying anti-HIV-1 genes were similar to those transduced by the control LN vectors. Stromal cells cultured from marrow samples from HIV-1-infected donors showed similar growth kinetics, hematopoietic support function, and enhancement of retrovirus-mediated transduction of CD34+ cells as seen with stromal cells cultured from uninfected marrow donors. Semi-quantitative polymerase chain reaction (PCR) was performed before and after ex vivo transduction to determine the frequency of HIV-1-containing cells in the CD34+ cell preparations. Although HIV-1+ cells were present at low levels in the mononuclear cell fractions of some of the marrow samples, the CD34+ cell preparation from only one marrow sample contained detectable HIV-1 positive cells (< 1 positive cell/100,000 by PCR) prior to transduction. None of the CD34+ cell preparations contained detectable HIV-1 after transduction. These studies demonstrate that HIV-1-infected patients are candidates for retrovirus-mediated transduction of anti-HIV-1 genes in bone marrow gene therapy clinical trials.

Original languageEnglish (US)
Pages (from-to)301-311
Number of pages11
JournalHuman Gene Therapy
Volume8
Issue number3
StatePublished - Feb 10 1997
Externally publishedYes

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Retroviridae
HIV-1
Bone Marrow
Tissue Donors
Genes
Stem Cells
CD4 Lymphocyte Count
Stromal Cells
Colony-Forming Units Assay
Polymerase Chain Reaction
Methylcellulose

ASJC Scopus subject areas

  • Genetics

Cite this

Kearns, K., Bahner, I., Bauer, G., Wei, S. F., Valdez, P., Wheeler, S., ... Kohn, D. B. (1997). Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer. Human Gene Therapy, 8(3), 301-311.

Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer. / Kearns, Karen; Bahner, Ingrid; Bauer, Gerhard; Wei, Sui Fang; Valdez, Penelope; Wheeler, Sarah; Woods, Lin; Miller, Robert; Casciato, Dennis; Galpin, Jeffrey; Church, Joseph; Kohn, Donald B.

In: Human Gene Therapy, Vol. 8, No. 3, 10.02.1997, p. 301-311.

Research output: Contribution to journalArticle

Kearns, K, Bahner, I, Bauer, G, Wei, SF, Valdez, P, Wheeler, S, Woods, L, Miller, R, Casciato, D, Galpin, J, Church, J & Kohn, DB 1997, 'Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer', Human Gene Therapy, vol. 8, no. 3, pp. 301-311.
Kearns K, Bahner I, Bauer G, Wei SF, Valdez P, Wheeler S et al. Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer. Human Gene Therapy. 1997 Feb 10;8(3):301-311.
Kearns, Karen ; Bahner, Ingrid ; Bauer, Gerhard ; Wei, Sui Fang ; Valdez, Penelope ; Wheeler, Sarah ; Woods, Lin ; Miller, Robert ; Casciato, Dennis ; Galpin, Jeffrey ; Church, Joseph ; Kohn, Donald B. / Suitability of bone marrow from HIV-1-infected donors for retrovirus-mediated gene transfer. In: Human Gene Therapy. 1997 ; Vol. 8, No. 3. pp. 301-311.
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