Bone marrow extracellular matrix molecules improve gene transfer into human hematopoietic cells via retroviral vectors.

Möritz, Thomas; Patel, Vikram; Williams, David A.

doi:10.1172/jci117122

Cited by 220 publications

(92 citation statements)

References 54 publications

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“…The combination of SCF + Flt3-L + Tpo resulted in transduction rates statistically indistinguishable from the standard combination. Consistent with previous reports for marrow from mice, 24 humans, 23,24,39 and baboons, 12 we observed more than a doubling in the transduction rates when we carried out transductions with the standard cytokine combination in plastic dishes coated with CH-296, with an average of 51.5 ± 2.0% G418-resistant colonies observed under these conditions in five separate experiments (compared with 24.7 ± 3.8% for control transductions carried out on untreated tissue culture plastic).…”

Section: Effect Of Culture Additives On Transduction Ratessupporting

confidence: 93%

“…Consistent with studies using marrow from mice, 24 humans, 23,24,39 and baboons, 12 our results demonstrate that coating culture vessels with CH-296 approximately doubles the rate of transduction for progenitors from pigtailed macaques. Because we only assessed the effect of CH-296 at 33°C, it is not clear whether the advantage gained by using CH-296-coated dishes would also be observed at 37°C.…”

Section: Figure 6 Effect Of Culture Length On Transduction Of Pigtailsupporting

confidence: 89%

“…38 In addition, we tried using culture vessels coated with the transduction-enhancing fibronectin fragment CH-296. 12, 23,39 As seen in Figure 5, combining Flt3-L alone or in combination with Tpo to the standard combination of IL-3 + IL-6 + SCF increased transduction rates 26% and 38% over the standard conditions alone (P р 0.001 each), while addition of IL-11 to the standard cytokine combination had no statistically significant effect. Combining IL-3 alone with Flt3-L, Tpo, or IL-11 resulted in transduction rates statistically lower than that achieved with the standard cytokine combination, as did the combinations of SCF + Flt3-L and SCF + Flt3-L + IL-11.…”

Section: Effect Of Culture Additives On Transduction Ratesmentioning

confidence: 94%

“…In order to optimize progenitor transduction rates, we also tested several critical parameters effecting the transduction efficiency of CD34-enriched marrow progenitors from this and other species, including the temperature and days of culture, the use of cytokines, and the use of dishes coated with the fibronectin fragment CH-296. 23,24 …”

Section: Introductionmentioning

confidence: 99%

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Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors

2000

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Nonhuman primates are increasingly being used as models for pre-clinical assessment of retrovirus vector expression and function following stem and progenitor cell transduction. We compared the relative susceptibility of CD34 + marrow progenitors from four nonhuman primate species and humans to transduction with amphotropic pseudotyped retrovirus vectors containing the Neo gene. The rate of functional gene transfer was measured by colony formation under G418 selection. Marrow progenitors from pigtail macaques (Macaca nemestrina) were transduced at about twice the rate (19.1 ± 4.3%) as those from rhesus (11.2 ± 3.7%) and cynomolgus (7.6 ± 1.9%) macaques, baboons (7.8 ± 1.8%), and humans (9.6 ± 1.7%). Semiquan-

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Section: Effect Of Culture Additives On Transduction Ratessupporting

confidence: 93%

Section: Figure 6 Effect Of Culture Length On Transduction Of Pigtailsupporting

confidence: 89%

Section: Effect Of Culture Additives On Transduction Ratesmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

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Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors

2000

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“…At the indicated time-points, cells were removed from plates containing the immobilized antibodies and transduced on fibronectin-coated plates essentially as described by Moritz et al 48 The recombinant fibronectin fragment (CH-296) which contains the connecting segment, cell-binding domain, and heparin-binding domain, 49 was kindly provided by Takara Shuzo (Otsu Shiga, Japan). Non-tissue culture-treated 24-well plates were coated with fibronectin (8 g/cm 2 ) for 2 h at room temperature.…”

Section: Retroviral Transductionsmentioning

confidence: 99%

Lentivirus-mediated gene transfer in primary T cells is enhanced by a central DNA flap

et al. 2001

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Retroviral vectors have become the primary tool for gene delivery into hematopoietic cells, including T lymphocytes. Lentiviral vectors offer an advantage over Moloney murine leukemia virus (MuLV) vectors because of their ability to translocate across an intact nuclear membrane and integrate into the genome of nonproliferating cells. We have recently demonstrated that a central strand displacement event, controlled by the central polypurine tract (cPPT) and the central termination sequence (CTS), results in the formation of a central DNA flap which acts as a cis-determinant of HIV-1 genome nuclear import. Here, we show that insertion of this DNA determinant in a classical lentiviral vector resulted in a significantly higher level of transduction in acti-

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Heparin-binding domain of human fibronectin binds HIV-1 gp120/160 and reduces virus infectivity

et al. 1998

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In vitro experiments indicate that components of the host present in body fluids may prevent the attachment of human immunodeficiency virus type 1 (HIV-1) to target cells. Fibronectin (Fn), a dimeric 440-kDa extracellular matrix adhesion protein, is secreted by mesenchymal cells and assembled into insoluble matrices. Fn exerts important effects on cell growth and differentiation through a number of discrete functional domains. Several microorganisms are known to bind Fn. We show that, under physiological conditions, HIV-1 gp120 and gp160 are capable of binding plasma and cellular Fn as well as laminin and vitronectin. Experiments were set up to analyze in detail the binding of HIV gp120 and gp160 to Fn. The gp120 and gp160 specifically recognize the C-terminal heparin-binding domain of Fn (Fn-CTHBD) with a calculated KD of 2.8 x 10(-7) M for gp160. Binding of gp160 to Fn-CTHBD is a saturable and specific process that is blocked by antibodies to Fn-CTHBD and by heparin and is inhibited to a minor extent by heparan sulfate and dextran sulfate. These observations suggest that gp120/160 specifically recognize the III15 repeat within Fn-CTHBD. Intact Fn and Fn-CTHBD strongly inhibit the interaction of gp120/160 with soluble CD4 and, under low serum conditions, are capable of neutralizing the infectivity of HIV-1 for CD4-positive T cells. Thus, Fn that is present in plasma and mucinous secretions may well affect HIV infectivity and virus distribution in vivo.

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Bone marrow extracellular matrix molecules improve gene transfer into human hematopoietic cells via retroviral vectors.

Cited by 220 publications

References 54 publications

Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors

Differences among nonhuman primates in susceptibility to bone marrow progenitor transduction with retrovirus vectors

Lentivirus-mediated gene transfer in primary T cells is enhanced by a central DNA flap

Heparin-binding domain of human fibronectin binds HIV-1 gp120/160 and reduces virus infectivity

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