Translational and clinical immunologyEfficacy of lentivirus-mediated gene therapy in an Omenn syndrome recombination-activating gene 2 mouse model is not hindered by inflammation and immune dysregulation
Section snippets
LV production
The 2.6kbUCOE-RAG2co LV, in which the human codon-optimized RAG2 cDNA was driven by a 2.6kbUCOE, was produced as previously described.26 For the 2.2kb UCOE-RAG2co LV, the 2.6kbUCOE was replaced by a shorter 2.2kbUCOE form.28
Animals
Animal experimental procedures were approved by the Institutional Animal Care and Use Committee of San Raffaele Hospital and Italian Ministry of Health (Institutional Animal Care and Use Committee no. 710). C57Bl/6 wild-type (WT) mice were obtained from Charles River (Bar
GT improves peripheral B- and T-cell counts in mice with OS
Mice with OS were lethally irradiated and transplanted with Lin− cells that were previously transduced with LV 2.6kbUCOE-RAG2co or 2.2kbUCOE-RAG2co (GT mice) to study the efficacy of GT.26, 28 Additional mice were treated with untransduced WT or OS Lin− cells as positive (BMT WT mice) and negative (BMT OS mice) controls, respectively. Starting at 6 weeks after treatment, we observed the appearance of B lymphocytes in the peripheral blood of GT mice with both vectors in marked contrast to BMT OS
Discussion
Mutations in RAG genes are responsible for a broad spectrum of clinical manifestations. Allogeneic HSCT is the standard of care, ideally performed before onset of life-threatening complications and organ damage, but its overall outcome remains unsatisfactory and can be recommended only when a matched donor is available.12, 17, 18, 40 In this scenario GT might represent a powerful alternative approach, as demonstrated by encouraging results obtained in recent clinical primary immunodeficiency
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Gene Therapy for Inborn Errors of Immunity
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2023, Seminars in ImmunologyGene Therapy for Inborn Errors of Immunity: Severe Combined Immunodeficiencies
2022, Hematology/Oncology Clinics of North AmericaCitation Excerpt :Residual RAG function partially permits T cell development leading to immune dysregulation. The efficacy of lentiviral-mediated gene therapy was tested in a murine model of Omenn syndrome.64 There was significant amelioration of immunodeficiency in treated mice, with the normalization of immunoglobulin levels and T-cell repertoire, good response to in vivo challenges, and dramatic improvement of autoimmune manifestations.
Efficacy and safety of anti-CD45–saporin as conditioning agent for RAG deficiency
2021, Journal of Allergy and Clinical ImmunologyCitation Excerpt :The levels of the liver enzymes alanine aminotransferase and aspartate aminotransferase were measured as described in the Methods section of the Online Repository (at www.jacionline.org). In vivo challenge with the T-dependent antigen 2,4,6-trinitrophenyl (TNP)-conjugated keyhole limpet hemocyanin was performed 4 months after transplantation as previously described.26 TNP-specific antibody titers were measured in serum by ELISA (see the Methods section of the Online Repository).
Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells
2020, Journal of Allergy and Clinical ImmunologyCitation Excerpt :The TCR repertoire was evaluated by means of deep sequencing, and data were analyzed with R Studio software. Screening for IgM/IgG reactivity against autoantigens was performed by using autoantibody arrays.63,64 The detailed methods are available in this article's Online Repository at www.jacionline.org.
Severe combined immune deficiency
2020, Stiehm's Immune Deficiencies: Inborn Errors of Immunity
Supported by the Italian Telethon foundation (Telethon Core TGT E2 Grant to A.V.); by the European Commission's 5th, 6th, and 7th and Horizon 2020 Framework Programs (contracts QLK3-CT-2001-00427-INHERINET, LSHB-CT-2004-005242-CONSERT, grant agreement 261387 CELL-PID to A.V. and grant 666908-SCIDNET to A.V.); and by The Netherlands Health Research and Development Organization ZonMw, Translational Gene Therapy Program (project 43100016 to G.W.) and by Italian Ministry of Health (grant GR-2011-02349759 to B.C.).
Disclosure of potential conflict of interest: L. D. Notarangelo is Editor of Frontiers in Immunology, Associate Editor for the Journal of Clinical Immunology, and Associate Editor for Clinical Immunology; is employed by the National Institutes of Health and the Department of Health and Human Services; and has received royalties from UpToDate. G. Wagemaker has received grants and travel support from The Netherlands Organization for Health Research ZonMw and the European Commission. A. Villa has received a grant from the European Commission. The rest of the authors declare that they have no relevant conflict of interest.