Suppression of FVIII Inhibitor Formation in Hemophilic Mice by Delivery of Transgene Modified Apoptotic Fibroblasts

Su, Rui; Epp, Angela; Latchman, Yvette; Bolgiano, Doug; Pipe, Steven W.; Josephson, Neil C.

doi:10.1038/mt.2009.209

Cited by 16 publications

(23 citation statements)

References 48 publications

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“…28 Hemophilia A mice treated with hFVIII expression vector-modified apoptotic fibroblasts demonstrated antigen-specific suppression of the immune response to hFVIII. 28 In this current study, we took a direct approach to delivering hFVIII antigen to APCs by transducing ex vivo generated tDCs with a foamy virus vector that carries a bioengineered hFVIII transgene. We demonstrated that intravenous delivery of vectormodified tDCs suppressed the inhibitor and T cell responses against hFVIII in both naive (Figures 2 and 7) and preimmunized (Figure 3) mice.…”

Section: Discussionmentioning

confidence: 99%

“…T cell proliferation assays were carried out as previously described. 28,40 In brief, 4-5 × 10 5 irradiated (3000 rad) CD90 depleted splenocytes were cocultured in wells of 96 well plates with 1 × 10 5 splenic CD4 + CD25 − T cells from pools of 3-5 mice in 200 µl/well RPMI 1640 complete media supplemented with increasing concentrations of rhFVIII or OVA. After 72 hours of coculture, 1 µCi 3 H-thymidine was added to the media and 18 hours later, cells were harvested.…”

Section: Methodsmentioning

confidence: 99%

“…Anti-OVA IgG titers were determined by ELISA as previously described. 28,41 Real-time PCR. Viral vector copy number in transduced tDCs was determined by real-time PCR using a DNA standard curve generated from serial dilutions of cells that contained a single copy of a foamy virus vector into nontransduced cells.…”

Section: Foamy Virus Vectorsmentioning

confidence: 99%

“…The FVIII inhibitor titer was determined by a standard Bethesda assay with minor modifications as previously described 28,42 Adoptive transfer studies. CD4 + cells from the spleens of donor mice were isolated at 90 hours after the last dose of three weekly infusions of tDCs.…”

Section: Foamy Virus Vectorsmentioning

confidence: 99%

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Suppression of the Immune Response to FVIII in Hemophilia A Mice by Transgene Modified Tolerogenic Dendritic Cells

Epp

Feng

et al. 2011

Molecular Therapy

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Current methods for eradicating clinically significant inhibitory antibodies to human factor VIII (hFVIII) in patients with hemophilia A rely on repeated delivery of high doses of factor concentrates for a minimum of many months. We hypothesize that tolerance can be induced more efficiently and reliably through hFVIII antigen presentation by tolerogenic dendritic cells (tDCs). In this study, we generated tDCs from hemophilia A mice and modified them with a foamy virus vector expressing a bioengineered hFVIII transgene. Naive and preimmunized mice infused with hFVIII expressing tDCs showed suppression of the T cell and inhibitor responses to recombinant hFVIII (rhFVIII). Treatment with hFVIII expressing tDCs was also associated with a higher percentage of splenocytes demonstrating a regulatory T cell phenotype in immunized mice. Furthermore, CD4(+) T cells harvested from recipients of hFVIII expression vector-modified tDCs were able to mediate antigen-specific immune suppression in naive secondary recipients. We also demonstrated a trend for improved suppression of inhibitor formation by coexpressing interleukin-10 (IL-10) and hFVIII from a bicistronic vector. These preclinical results demonstrate the potential for employing vector modified ex vivo generated tDCs to treat high titer inhibitors in patients with hemophilia A.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Foamy Virus Vectorsmentioning

confidence: 99%

Section: Foamy Virus Vectorsmentioning

confidence: 99%

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Suppression of the Immune Response to FVIII in Hemophilia A Mice by Transgene Modified Tolerogenic Dendritic Cells

Epp

Feng

et al. 2011

Molecular Therapy

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“…demonstrated that injection of fVIII expression vector-modified apoptotic syngeneic fibroblasts achieves suppression of fVIII inhibitor development in hemophilia A mice. 12 However, the significant issues in this approach are the complex procedures such as collection of fibroblasts from each patient and transfection of the F8 gene into these cells. Therefore, fVIII-expressing cells derived from histocompatible stem cells [such as embryonic stem (ES) cells or induced pluripotent stem (iPS) cells] will be the most suitable.…”

Section: Introductionmentioning

confidence: 99%

Repression of Factor VIII Inhibitor Development with Apoptotic Factor VIII-Expressing Embryonic Stem Cells

et al. 2013

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Development of factor VIII (fVIII)-neutralizing antibodies, called inhibitors, is a challenging problem in the management of hemophilia A patients. We explored the possibility of pretreatment with apoptotic fVIII-expressing embryonic stem (ES) cells to prevent the development of fVIII inhibitors. Murine ES cells integrated with the human F8 gene were differentiated into embryoid bodies, dissociated to a single cell suspension, subjected to hypo-osmotic shock to induce apoptosis, and intraperitoneally injected into hemophilia A mice. Inhibitors were induced by periodic intraperitoneal injections of recombinant human fVIII (rhfVIII). In the groups in which intraperitoneal injections of rhfVIII began at 1-3 weeks after pretreatment, the titers of inhibitors were significantly lower after the third administration of rhfVIII compared with that in the control group in which apoptotic Ainv18 ES cells (without the human F8 gene) were used for pretreatment, and continued to show lower levels until the sixth administration of rhfVIII. These results suggest that pretreatment with apoptotic hfVIII-expressing ES cells might be promising for the prevention of fVIII inhibitor development in hemophilia A patients.

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Synthesis and assembly of dengue virus envelope protein fused to cholera toxin B subunit into biologically active oligomers in transgenic tomato (Solanum lycopersicum)

Kim

Yang

2016

Plant Biotechnol Rep

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Dengue is the fastest growing mosquito-borne disease worldwide, causing nearly 400 million infections annually. A universally applicable dengue virus vaccine is required to arrest its spread. Here, we generated an edible dengue vaccine by expressing the dengue fusion protein in tomatoes, which is a desirable expression system owing to the inherent adjuvanticity of alpha tomatine and immunogenicity of the tomato lectin/microbial antigen complex. The B subunit of Vibrio cholera toxin (CTB) was genetically fused to dengue envelope antigen for improved delivery to antigen-presenting cells and enhanced immunogenicity, while avoiding immunological tolerance. We utilized domain III of the dengue envelope protein (EDIII), as it has been shown to induce serotype-specific neutralizing antibodies. The CTB-EDIII fusion gene construct containing an endoplasmic reticulum target sequence was introduced into tomato plants by Agrobacterium tumefaciens-mediated gene transformation, and the expression of CTB-EDIII in transgenic plants was confirmed by DNA, RNA and protein analyses. Accumulated fusion protein accounted for up to 0.015 % of total soluble protein, and it assembled into fully functional pentamers as demonstrated by binding to GM1 ganglioside. Future work will involve testing of transgenic tomatoes for immunogenicity in mice following oral delivery.

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Suppression of FVIII Inhibitor Formation in Hemophilic Mice by Delivery of Transgene Modified Apoptotic Fibroblasts

Cited by 16 publications

References 48 publications

Suppression of the Immune Response to FVIII in Hemophilia A Mice by Transgene Modified Tolerogenic Dendritic Cells

Suppression of the Immune Response to FVIII in Hemophilia A Mice by Transgene Modified Tolerogenic Dendritic Cells

Repression of Factor VIII Inhibitor Development with Apoptotic Factor VIII-Expressing Embryonic Stem Cells

Synthesis and assembly of dengue virus envelope protein fused to cholera toxin B subunit into biologically active oligomers in transgenic tomato (Solanum lycopersicum)

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