Gene therapy to inhibit xenoantibody production using lentiviral vectors in non-human primates

Fischer-Lougheed, Jacqueline; Tarantal, Alice F.; Shulkin, Irina; Mitsuhashi, Noboru; Kohn, Donald B.; Lee, Charles C.; Kearns‐Jonker, Mary

doi:10.1038/sj.gt.3302818

Cited by 12 publications

(12 citation statements)

References 42 publications

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“…In order to determine whether induced anti-non-gal xenoantibodies can bind to carbohydrate xenoantigens, we transduced GalTKO pig cells with lentiviral vectors expressing porcine α1,3 galactosyltransferase (pCSO-rre-cppt-MCU3-α1,3gal-WPRE) or a control lentivector (αGT in reverse orientation) 30 . The GalTKO pig cells were plated at 1×10 5 cells per well and were transduced in 0.5 ml of Medium 199 (Invitrogen, Carlsbad, CA).…”

Section: Methodsmentioning

confidence: 99%

The Anti-Non-Gal Xenoantibody Response to Xenoantigens on Gal Knockout Pig Cells Is Encoded by a Restricted Number of Germline Progenitors

Kiernan

Harnden

Gunthart

et al. 2008

American Journal of Transplantation

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Antibodies directed at non-gal xenoantigens are responsible for acute humoral xenograft rejection when gal knockout (GalTKO) pig organs are transplanted into non-human primates. We generated IgM and IgG gene libraries using peripheral blood lymphocytes of rhesus monkeys initiating active xenoantibody responses after immunization with GalTKO pig endothelial cells and used these libraries to identify IgVH genes that encode antibody responses to non-gal pig xenoantigens. Immunoglobulin genes derived from the IGHV3-21 germline progenitor encode xenoantibodies directed at non-gal xenoantigens. Transduction of GalTKO cells with lentiviral vectors expressing the porcine α1,3 galactosyltransferase gene responsible for gal carbohydrate expression results in a higher level of binding of “anti-non-gal” xenoantibodies to transduced GalTKO cells expressing the gal carbohydrate, suggesting that anti-non-gal xenoantibodies crossreact with carbohydrate xenoantigens. The galactosyltransferase 2 gene encoding isoglobotriaosylceramide synthase (iGb3 synthase) is not expressed in GalTKO pig cells. Our results demonstrate that anti-non-gal xenoantibodies in primates are encoded by IgVH genes that are restricted to IGHV3-21 and bind to an epitope that is structurally related to but distinct from the Gal carbohydrate.

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Section: Methodsmentioning

confidence: 99%

The Anti-Non-Gal Xenoantibody Response to Xenoantigens on Gal Knockout Pig Cells Is Encoded by a Restricted Number of Germline Progenitors

Kiernan

Harnden

Gunthart

et al. 2008

American Journal of Transplantation

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“…Anti-aGal Abs were not produced in GalT-reconstituted mice, in contrast to anti-non-Gal xenoAb. Fischer-Lougheed et al 17 reproduced these results in rhesus macaques: several months post-BM transplantation, monkeys were immunized with porcine cells, and the anti-aGal IgM induction was considerably attenuated in aGal+BM recipients compared to immunized controls (no transplantation was performed at this stage). In these studies, the authors reported that this type of gene therapy approach was able to inhibit the production of anti-aGal Ab both in mice and in rhesus macaques, suggesting that B-cell tolerance achieved in mice could also be achieved in NHP.…”

Section: Genetic Engineering For Molecules That Modify the Cellular Xmentioning

confidence: 87%

“…17 Mitsuhashi et al 16 used lentiviral transfer of the GalT gene to autologous BM from GalT-KO mice and then transplanted the BM cells into submyeloablative irradiated GalT-KO mice. Following BM transplantation, the mice were immunized with rabbit red blood cells and subsequently grafted with aGal+ mice hearts.…”

Section: Genetic Engineering For Molecules That Modify the Cellular Xmentioning

confidence: 99%

Progress and prospects: genetic engineering in xenotransplantation

2008

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In this review, we summarize the work published over the last 2 years using genetic modifications of animals in the field of xenotransplantation. Genetic engineering of the donor has become a powerful tool in xenotransplantation, both for the inactivation of one particular porcine gene and for the addition of human genes with the goal of overcoming xenogeneic barriers. We summarize the work relative to the knockout of the a1,3-galactosyltransferase gene, followed by genetic engineering aimed at reducing the humoral and cellular immune response, complement activation and coagulation. Finally, we report on the genetic modification of pigs to reduce porcine endogenous retrovirus infection risk in the xenogeneic context.

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“…Our laboratory recently used SIV-based lentiviral vectors to achieve chimerism prior to immunization of rhesus monkeys with porcine hepatocytes [Fischer-Lougheed et al, 2007]. In this study, we reported that GalT-encoding lentiviral vectors can be used to achieve chimerism sufficient to inhibit xenoantibody production in non-human primates after sublethal irradiation.…”

Section: Gene Therapy For Xenotransplantation In Non-human Primatesmentioning

confidence: 93%

Gene Therapy for the Induction of Chimerism and Transplant Tolerance

Gunthart¹,

Kearns‐Jonker²

2007

CGT

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Technical advances in transplant surgery and the development of powerful and effective immunosuppressive drugs have contributed to the success of organ transplantation as a medical treatment for patients with end-stage diseases. Associated with this procedure, however, is a dependence on life-long immunosuppressive drugs, which are required to prevent graft rejection. These agents render the patient susceptible to infections, tumors and various side affects. The ability to achieve tolerance to organ grafts would free transplant patients from lifelong dependency on pharmacological agents with harmful side effects. Several laboratories have shown that tolerance can be achieved by the induction of mixed cell chimerism and/or by molecular chimerism achieved by gene transfer techniques prior to graft placement. Molecular chimerism, induced by transplantation of autologous bone marrow expressing either allo- or xenoantigens has the potential to induce tolerance without the development of graft vs. host disease. The application of gene transfer techniques to induce chimerism has been shown to reshape the immune repertoire by mechanisms that include clonal deletion, the induction of central tolerance or generation of regulatory T cells that would eliminate the need for immunosuppressive drugs. Optimization of this methodology for clinical use could therefore revolutionize the field of transplantation. This review summarizes the recent studies which have compared the efficacy of different vectors, conditioning regimens, and transduction conditions leading to new and improved techniques for the application of gene therapy to induce chimerism and transplant tolerance to both allografts and xenografts.

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Gene therapy to inhibit xenoantibody production using lentiviral vectors in non-human primates

Cited by 12 publications

References 42 publications

The Anti-Non-Gal Xenoantibody Response to Xenoantigens on Gal Knockout Pig Cells Is Encoded by a Restricted Number of Germline Progenitors

The Anti-Non-Gal Xenoantibody Response to Xenoantigens on Gal Knockout Pig Cells Is Encoded by a Restricted Number of Germline Progenitors

Progress and prospects: genetic engineering in xenotransplantation

Gene Therapy for the Induction of Chimerism and Transplant Tolerance

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