Intracellular immunization of human fetal cord blood stem/progenitor cells with a ribozyme against human immunodeficiency virus type 1.

Yu, Meichen; Leavitt, Mark C.; Maruyama, Midori; Yamada, Osamu; Young, Dennis; Ho, Anthony D.; Wong‐Staal, Flossie

doi:10.1073/pnas.92.3.699

Cited by 104 publications

(66 citation statements)

References 25 publications

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“…Delivery of anti-HIV ribozyme, dominant negative proteins, or decoys to CD34+ cells partially protected progeny MDMs from HIV replication. 47,[49][50][51][52] Recently, lentivirus vectors used to deliver RNAi against CCR5 were reported to transduce PBMC with low efficiency, and partially protect them from HIV replication. 53 We have found that rSV40 delivery of RNAi may be highly effective in protecting MDMs from R5-tropic HIV.…”

Section: Discussionmentioning

confidence: 99%

Protecting from R5-tropic HIV: individual and combined effectiveness of a hammerhead ribozyme and a single-chain Fv antibody that targets CCR5

Cordelier

Kulkowsky

et al. 2004

Gene Ther

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The CCR5 chemokine receptor is important for most clinical strains of HIV to establish infection. Individuals with naturally occurring polymorphisms in the CCR5 gene who have reduced or absent CCR5 are apparently otherwise healthy, but are resistant to HIV infection. With the goal of reducing CCR5 and protecting CCR5+ cells from R5-tropic HIV, we used Tag-deleted SV40-derived vectors to deliver several anti-CCR5 transgenes: 2C7, a single-chain Fv (SFv) antibody; VCKA1, a hammerhead ribozyme; and two natural CCR5 ligands, MIP-1a and MIP-1b, modified to direct these chemokines, and hence their receptor to the endoplasmic reticulum. These transgenes were delivered using recombinant, Tag-deleted SV40-derived vectors to human CCR5+ cell lines and primary cells: monocyte-derived macrophages and brain microglia. All transgenes except MIP-1a decreased CCR5, as assayed by immunostaining, Northern blotting, and cytofluorimetry (FACS). Individually, all transgenes except MIP-1a protected from low challenge doses of HIV. At higher dose HIV challenges, protection provided by all transgenes diminished, the SFv and the ribozyme being most potent. Vectors carrying these two transgenes were used sequentially to deliver combination anti-CCR5 genetic therapy. This approach gave approximately additive reduction in CCR5, as measured by FACS and protected from higher dose HIV challenges. Reducing cell membrane CCR5 using anti-CCR5 transgenes, alone or in combinations, may therefore provide a degree of protection from R5-tropic strains of HIV.

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Section: Discussionmentioning

confidence: 99%

Protecting from R5-tropic HIV: individual and combined effectiveness of a hammerhead ribozyme and a single-chain Fv antibody that targets CCR5

Cordelier

Kulkowsky

et al. 2004

Gene Ther

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“…In vitro studies on PUCB CD34 + cells have indicated that retrovirus-transduced genes can be introduced into 12-95% of progenitor cells. 9,[11][12][13] In vivo animal studies using SCID mouse models have demonstrated that CD34 + cells derived from PUCB could be successfully engrafted. 14 These promising data have recently led to the initiation of a human gene therapy trial using PUCB-derived CD34 + cells for adenosine deaminase deficiency.…”

Section: Discussionmentioning

confidence: 99%

“…9 Colony formation of Table 3 Clonogenic formation of transduced CD34 + cells derived from normal, HIV-1-exposed and HIV-1-infected infants…”

Section: Ribozyme Transduction and Expression In Transduced Cellsmentioning

confidence: 99%

“…15 Previous studies from many groups have demonstrated the feasibility of transducing CD34 + cells from normal PUCB samples. 9 However, it was not known if CD34 + cells from HIV-1-exposed or HIV-1-infected PUCB samples were functionally intact and could be successfully transduced, and if the transgene would be functional in the progeny cells. It was also not clear if CD34 + cells from such examples were themselves infected by HIV-1 and if such cells could be used as vehicles for gene therapy.…”

Section: Figure 3 Hiv-1 Challenge Of Ribozyme Transduced Cells Macromentioning

confidence: 99%

“…Moreover, macrophage progeny of the transduced CD34 + cells was resistant to challenges from monocyte-tropic HIV-1. 9 Although it is conceptually feasible to use transduced stem cells in an allogeneic setting, the need for myeloablation and post-infusion immunological suppressions, such as treatment of graft-versus-host disease, make it impractical for a phase I clinical trial. Therefore, it is more logical to use PUCB CD34 + cells from an autologous source initially to test the concept of gene therapy for HIV-1 infection.…”

Section: Introductionmentioning

confidence: 99%

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Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

Gervaix

Kang

et al. 1998

Gene Ther

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Hematopoietic CD34+ cells from placental and umbilical cord blood (PUCB) can be valuable vehicles for gene therapy of immunodeficiencies and genetic disorders. We have conducted preclinical studies towards the treatment of HIV-1-infected infants with genetically 'immunized' CD34+ cells derived from PUCB using anti-HIV-1 hairpin ribozyme genes. PUCB was collected from 10 newborns of HIV-1-positive mothers. CD34+ cells were enriched with a modified procedure using Dynal immunomagnetic beads and chymopapain, stimulated with stem cell factor (SCF), IL-3 and IL-6, and transduced using cell-free recombinant retroviral vector (MJT) expressing a ribozyme against the U5 region of HIV-1. No significant differences were observed in the growth pattern of CD34+ cells from normal infants, HIV-1 exposed infants or infants confirmed to be infected by HIV-1. The transduction efficiency of the CD34+ cells from all the infants was also comparable. MJT-transduced CD34+ cells from an HIV-1-infected infant were maintained in a liquid culture system for 4 weeks, and the progeny macrophage cells were challenged with the monocyte-tropic laboratory strain, HIV-Bal, or the HIV-1 isolate from the infant's mother. Significant inhibition of virus replication was observed in ribozyme-transduced cells. Thus, we have demonstrated efficient and stable gene transfer into progenitor cells from the cord blood of HIV-1-exposed or -infected infants and shown that protection from HIV-1 infection was conferred to the progeny cells produced by CD34+ cells transduced with the anti-HIV ribozyme gene construct

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Retroviral gene transfer into cord blood stem/progenitor cells using purified vector stocks

et al. 1998

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Cord blood (CB) progenitor/stem cells (P/SC) are ideal targets for early gene therapy in individuals prenatally diagnosed with genetic disorders. Most retroviral transduction protocols were developed using adult peripheral blood stem cells (PBSC) and bone marrow (BM). Less is known about retroviral transduction of CB P/SC. We examined how timing, multiplicity of infection (MOI), and polycations in the transduction media affect transduction efficiency. Rates of transduction were determined in recently isolated CD34+ enriched CB cells and in colonies derived after various times in liquid cultures (LC). CB mononuclear cells (MNC) were separated by ficoll-hypaque centrifugation and enriched for CD34+ cells. Purity was assessed by flow cytometry. Transduction were performed with clinical-grade retroviral stocks at MOIs of 1-20. Transduction was performed with fetal bovine serum (FBS) or autologous plasma, IL-3, GM-CSF, IL-6, and SCF. The retroviral vector contained LacZ and neomycin resistance (neo) reporter genes. Transduction was determined by X-gal stain and by PCR amplification of the reporter genes. No drug selection was used. Twenty-five experiments were done. CB volumes ranged from 35-150 ml. MNC and CD34+ cell counts ranges were: 0.14-840 x 10(6) and 0.1-4.2 x 10(6), respectively. Transduction efficiency in liquid cultures ranged from 4-63%. Higher rates were seen using MOI > or = 10, 2 microg/ml polybrene, and 10% autologous CB plasma. In colonies, transduction rates were 63 to 72% by PCR and 32% by X-gal staining. In LTC-IC derived colonies, transduction was 7% by PCR. Short incubations of CD34+ CB cells with purified retroviral stocks, polybrene, and autologous sera result in high transduction rates of committed progenitors and moderately low efficiencies of transduction of LTC-IC in the absence of drug selection.

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Intracellular immunization of human fetal cord blood stem/progenitor cells with a ribozyme against human immunodeficiency virus type 1.

Cited by 104 publications

References 25 publications

Protecting from R5-tropic HIV: individual and combined effectiveness of a hammerhead ribozyme and a single-chain Fv antibody that targets CCR5

Protecting from R5-tropic HIV: individual and combined effectiveness of a hammerhead ribozyme and a single-chain Fv antibody that targets CCR5

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

Retroviral gene transfer into cord blood stem/progenitor cells using purified vector stocks

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