6 results on '"Hernandez RJ"'
Search Results
2. Lentiviral Vector Gene Therapy Protects XCGD Mice From Acute Staphylococcus aureus Pneumonia and Inflammatory Response
- Author
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Bernhard Gentner, Chiara Bovolenta, Clelia Di Serio, Maddalena Migliavacca, Raisa Jofra Hernandez, Alice Rossi, Alessandra Bragonzi, Serena Ranucci, Francesca Sanvito, Aleksandar Pramov, Chiara Brombin, Giada Farinelli, Alessandro Aiuti, Farinelli, G, Hernandez, Rj, Rossi, A, Ranucci, S, Sanvito, F, Migliavacca, M, Brombin, Chiara, Pramov, A, DI SERIO, Mariaclelia, Bovolenta, C, Gentner, B, Bragonzi, A, and Aiuti, Alessandro
- Subjects
0301 basic medicine ,Staphylococcus aureus ,Chemokine ,Genetic enhancement ,Genetic Vectors ,Inflammation ,Granulomatous Disease, Chronic ,medicine.disease_cause ,Viral vector ,Proinflammatory cytokine ,Mice ,03 medical and health sciences ,Chronic granulomatous disease ,Pneumonia, Staphylococcal ,Drug Discovery ,Genetics ,medicine ,Animals ,Humans ,Vector (molecular biology) ,Molecular Biology ,Cells, Cultured ,Pharmacology ,Membrane Glycoproteins ,biology ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,NADPH Oxidases ,Genetic Therapy ,Hematopoietic Stem Cells ,medicine.disease ,Bacterial Load ,3. Good health ,Disease Models, Animal ,030104 developmental biology ,NADPH Oxidase 2 ,Immunology ,biology.protein ,Cytokines ,Molecular Medicine ,Original Article ,Chemokines ,medicine.symptom - Abstract
Chronic granulomatous disease (CGD) is a primary immunodeficiency due to a deficiency in one of the subunits of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. CGD patients are characterized by an increased susceptibility to bacterial and fungal infections, and to granuloma formation due to the excessive inflammatory responses. Several gene therapy approaches with lentiviral vectors have been proposed but there is a lack of in vivo data on the ability to control infections and inflammation. We set up a mouse model of acute infection that closely mimic the airway infection in CGD patients. It involved an intratracheal injection of a methicillin-sensitive reference strain of S. aureus . Gene therapy, with hematopoietic stem cells transduced with regulated lentiviral vectors, restored the functional activity of NADPH oxidase complex (with 20–98% of dihydrorhodamine positive granulocytes and monocytes) and saved mice from death caused by S. aureus , significantly reducing the bacterial load and lung damage, similarly to WT mice even at low vector copy number. When challenged, gene therapy-treated XCGD mice showed correction of proinflammatory cytokines and chemokine imbalance at levels that were comparable to WT. Examined together, our results support the clinical development of gene therapy protocols using lentiviral vectors for the protection against infections and inflammation.
- Published
- 2016
3. Ex vivo gene therapy with lentiviral vectors rescues adenosine deaminase (ADA)–deficient mice and corrects their immune and metabolic defects
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Filippo Carlucci, Luca Biasco, Claudio Doglioni, Antonella Tabucchi, Claudio Bordignon, Maurilio Ponzoni, Alessandro Aiuti, Antonia Follenzi, Raisa Jofra Hernandez, Alessandra Mortellaro, Luigi Naldini, Francesca Sanvito, Clelia Di Serio, Matteo M. Guerrini, Maria Grazia Roncarolo, Mortellaro, A, Hernandez, Rj, Guerrini, Mm, Carlucci, F, Tabucchi, A, Ponzoni, Maurilio, Sanvito, F, Doglioni, Claudio, DI SERIO, Mariaclelia, Biasco, L, Follenzi, A, Naldini, Luigi, Bordignon, Claudio, Roncarolo, MARIA GRAZIA, and Aiuti, Alessandro
- Subjects
Adenosine Deaminase ,Genetic enhancement ,Genetic Vectors ,Immunology ,Mice, Transgenic ,Lymphocyte Activation ,Biochemistry ,Viral vector ,Mice ,Adenosine deaminase ,Immune system ,medicine ,Animals ,Lymphocyte Count ,Bone Marrow Transplantation ,Mice, Knockout ,B-Lymphocytes ,Severe combined immunodeficiency ,biology ,Lentivirus ,Gene Transfer Techniques ,Cell Biology ,Hematology ,Flow Cytometry ,medicine.disease ,Adenosine deaminase deficiency ,Killer Cells, Natural ,Transplantation ,medicine.anatomical_structure ,Antibody Formation ,biology.protein ,Bone marrow ,Spleen - Abstract
Adenosine deaminase (ADA) deficiency is caused by a purine metabolic dysfunction, leading to severe combined immunodeficiency (SCID) and multiple organ damage. To investigate the efficacy of ex vivo gene therapy with self-inactivating lentiviral vectors (LVs) in correcting this complex phenotype, we used an ADA(-/-) mouse model characterized by early postnatal lethality. LV-mediated ADA gene transfer into bone marrow cells combined with low-dose irradiation rescued mice from lethality and restored their growth, as did transplantation of wild-type bone marrow. Mixed chimerism with multilineage engraftment of transduced cells was detected in the long term in animals that underwent transplantation. ADA activity was normalized in lymphocytes and partially corrected in red blood cells (RBCs), resulting in full metabolic detoxification and prevention of severe pulmonary insufficiency. Moreover, gene therapy restored normal lymphoid differentiation and immune functions, including antigen-specific antibody production. Similar degrees of detoxification and immune reconstitution were obtained in mice treated early after birth or after 1 month of enzyme-replacement therapy, mimicking 2 potential applications for ADA-SCID. Overall, this study demonstrates the efficacy of LV gene transfer in correcting both the immunological and metabolic phenotypes of ADA-SCID and supports the future clinical use of this approach. Adenosine deaminase (ADA) deficiency is caused by a purine metabolic dysfunction, leading to severe combined immunodeficiency (SCID) and multiple organ damage. To investigate the efficacy of ex vivo gene therapy with self-inactivating lentiviral vectors (LVs) in correcting this complex phenotype, we used an ADA(-/-) mouse model characterized by early postnatal lethality. LV-mediated ADA gene transfer into bone marrow cells combined with low-dose irradiation rescued mice from lethality and restored their growth, as did transplantation of wild-type bone marrow. Mixed chimerism with multilineage engraftment of transduced cells was detected in the long term in animals that underwent transplantation. ADA activity was normalized in lymphocytes and partially corrected in red blood cells (RBCs), resulting in full metabolic detoxification and prevention of severe pulmonary insufficiency. Moreover, gene therapy restored normal lymphoid differentiation and immune functions, including antigen-specific antibody production. Similar degrees of detoxification and immune reconstitution were obtained in mice treated early after birth or after 1 month of enzyme-replacement therapy, mimicking 2 potential applications for ADA-SCID. Overall, this study demonstrates the efficacy of LV gene transfer in correcting both the immunological and metabolic phenotypes of ADA-SCID and supports the future clinical use of this approach.
- Published
- 2006
4. IL-3 or IL-7 Increases ex Vivo Gene Transfer Efficiency in ADA-SCID BM CD34+ Cells while Maintaining in Vivo Lymphoid Potential
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Nicole Carballido-Perrig, Maria Grazia Roncarolo, Grazia Andolfi, Alessandro Aiuti, Augusto Colombo, Claudio Bordignon, Daniela Superchi, Raisa Jofra Hernandez, Cristina Mocchetti, José M. Carballido, Francesca Ficara, Sara Deola, Ficara, F, Superchi, Db, Hernandez, Rj, Mocchetti, C, Carballido Perrig, N, Andolfi, G, Deola, S, Colombo, A, Bordignon, Claudio, Carballido, Jm, Roncarolo, MARIA GRAZIA, and Aiuti, Alessandro
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Adenosine Deaminase ,Genetic enhancement ,T cell ,Antigens, CD34 ,Bone Marrow Cells ,Mice, SCID ,Biology ,Mice ,Transduction, Genetic ,In vivo ,Drug Discovery ,Genetics ,medicine ,Animals ,Humans ,Lymphocytes ,Progenitor cell ,Molecular Biology ,Cells, Cultured ,B cell ,Cell Proliferation ,Interleukin 3 ,Pharmacology ,Interleukin-7 ,Gene Transfer Techniques ,Cell Differentiation ,Genetic Therapy ,Fetal Blood ,Molecular biology ,medicine.anatomical_structure ,Immunology ,Molecular Medicine ,Interleukin-3 ,Severe Combined Immunodeficiency ,Bone marrow ,Ex vivo ,Stem Cell Transplantation - Abstract
To improve maintenance and gene transfer of human lymphoid progenitors for clinical use in gene therapy of adenosine deaminase (ADA)-deficient SCID we investigated several gene transfer protocols using various stem cell-enriched sources. The lymphoid differentiation potential was measured by an in vitro clonal assay for B/NK cells and in the in vivo SCID-hu mouse model. Ex vivo culture with the cytokines TPO, FLT3-ligand, and SCF (T/F/S) plus IL-3 or IL-7 substantially increased the yield of transduced bone marrow (BM) CD34(+) cells purified from ADA-SCID patients or healthy donors, compared to T/F/S alone. Moreover, the use of IL-3 or IL-7 significantly improved the maintenance of in vitro B cell progenitors from ADA-SCID BM cells and allowed the efficient transduction of B and NK cell progenitors. Under these optimized conditions transduced CD34(+) cells were efficiently engrafted into SCID-hu mice and gave rise to B and T cell progeny, demonstrating the maintenance of in vivo lymphoid reconstitution capacity. The protocol based on the T/F/S + IL-3 combination was included in a gene therapy clinical trial for ADA-SCID, resulting in long-term engraftment of stem/progenitor cells. Remarkably, gene-corrected BM CD34(+) cells obtained from one patient 4 and 11 months after gene therapy were capable of repopulating the lymphoid compartment of SCID-hu hosts.
- Published
- 2004
5. Efficacy of Gene Therapy for Wiskott-Aldrich Syndrome Using a WAS Promoter/cDNA-Containing Lentiviral Vector and Nonlethal Irradiation
- Author
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Loëc Dupré, Sara Trifari, Luigi Naldini, Samantha Scaramuzza, Raisa Jofra Hernandez, Maria Grazia Roncarolo, Francesco Marangoni, Alessandro Aiuti, Dupre, L, Marangoni, F, Scaramuzza, S, Trifari, S, Hernandez, Rj, Aiuti, Alessandro, Naldini, Luigi, and Roncarolo, MARIA GRAZIA
- Subjects
DNA, Complementary ,Wiskott–Aldrich syndrome ,T cell ,Genetic enhancement ,T-Lymphocytes ,Blotting, Western ,Genetic Vectors ,macromolecular substances ,Biology ,Transplantation, Autologous ,Viral vector ,Mice ,Transduction, Genetic ,medicine ,Genetics ,Animals ,Humans ,Fluorescent Antibody Technique, Indirect ,Promoter Regions, Genetic ,Molecular Biology ,Cell Proliferation ,B-Lymphocytes ,Reverse Transcriptase Polymerase Chain Reaction ,Lentivirus ,Hematopoietic Stem Cell Transplantation ,Genetic Therapy ,medicine.disease ,Virology ,Actins ,Wiskott-Aldrich Syndrome ,Transplantation ,Mice, Inbred C57BL ,Haematopoiesis ,medicine.anatomical_structure ,Primary immunodeficiency ,Cancer research ,Interleukin-2 ,Molecular Medicine ,Stem cell ,Wiskott-Aldrich Syndrome Protein - Abstract
Wiskott-Aldrich syndrome (WAS) is a life-threatening X-linked primary immunodeficiency characterized by infections, hemorrhages, autoimmune disorders, and lymphomas. Transplantation of genetically corrected autologous hematopoietic stem cells (HSCs) could represent an alternative treatment to allogeneic HSC transplantation, the latter being often associated with severe complications. We used WAS(-/-) mice to test the efficacy of a gene therapy approach based on nonlethal irradiation followed by transplantation of WAS(-/-) HSCs transduced with lentiviral vectors encoding the WAS protein (WASP) from either the ubiquitous PGK promoter or the tissue-specific WAS promoter. The procedure resulted in significant levels of engraftment of WASP-expressing T cells, B cells, platelets, and myeloid cells. T cells harbored one or two vector copies and displayed partial to full correction of T cell receptor-driven interleukin- 2 production and proliferation. In addition, polymerization of F-actin and localization of WASP at the site of the immunological synapse were restored. The treatment was well tolerated and no pathology was detected by systematic blood analysis and autopsy. The efficacy of WAS gene transfer into HSCs, using the WAS promoter-containing lentiviral vector, combined with nonlethal irradiation provides a strong rationale for the development of gene therapy for WAS patients. Wiskott-Aldrich syndrome (WAS) is a life-threatening X-linked primary immunodeficiency characterized by infections, hemorrhages, autoimmune disorders, and lymphomas. Transplantation of genetically corrected autologous hematopoietic stem cells (HSCs) could represent an alternative treatment to allogeneic HSC transplantation, the latter being often associated with severe complications. We used WAS-/- mice to test the efficacy of a gene therapy approach based on nonlethal irradiation followed by transplantation of WAS-/- HSCs transduced with lentiviral vectors encoding the WAS protein (WASP) from either the ubiquitous PGK promoter or the tissue- specific WAS promoter. The procedure resulted in significant levels of engraftment of WASP-expressing T cells, B cells, platelets, and myeloid cells. T cells harbored one or two vector copies and displayed partial to full correction of T cell receptor-driven interleukin-2 production and proliferation. In addition, polymerization of F-actin and localization of WASP at the site of the immunological synapse were restored. The treatment was well tolerated and no pathology was detected by systematic blood analysis and autopsy. The efficacy of WAS gene transfer into HSCs, using the WAS promoter-containing lentiviral vector, combined with nonlethal irradiation provides a strong rationale for the development of gene therapy for WAS patients
- Published
- 2006
6. [Contribution to the use of methergin in obstetrics].
- Author
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LOPEZ HERNANDEZ RJ
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- Female, Humans, Pregnancy, Cardiovascular Agents, Ergot Alkaloids therapeutic use, Labor, Obstetric, Methylergonovine, Obstetrics, Oxytocics, Work
- Published
- 1952
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