Showing papers by "Transgene SA published in 1997"
TL;DR: Treatment regimens with immunosuppressive drugs or with monoclonal antibodies that block either the T cell receptor or costimulation pathways allow prolonged transgene expression and/or readministration of adenoviral vectors and transduction efficiencies may be increased by transiently inhibiting non-specific immune mechanisms that lead to the dramatic early clearance of the vectors.
155 citations
TL;DR: Growing evidence now suggests that some transgenes can be subject to a phenomenon akin to the variegation found in insects and plants, which can complicate the interpretation of some experiments using transgenic animals generated by microinjection.
120 citations
TL;DR: It is demonstrated that infection of the pulmonary airways is dependent on the developmental stage of the foetus and can be achieved on the 15th day of gestation, and retrovirus producer cells could become a means to overcome the limitations of low retroviral titre for in vivo foetal gene transfer.
Abstract: With the aim of developing foetal gene therapy for cystic fibrosis, we have investigated the possibility of gene targeting to the mouse foetus with two different viral vector systems and at different times of gestation. We report here that recombinant retrovirus producing cells administered into the intra-amniotic cavity of mid- to late-gestation mouse MF1 foetuses survive in the amniotic fluid and are able to engraft to a certain extent in foetal tissues. By production of infectious virus they mediate transduction and beta-galactosidase transgene expression in neighbouring foetal tissues 24 to 72 h following injection. Retrovirus producer cells could, therefore, become a means to overcome the limitations of low retroviral titre, for in vivo foetal gene transfer. To investigate the developmental stage at which transduction of the airways and enteral systems can be obtained we also administered a highly infective first generation adenoviral vector (AdRSV beta gal) into the amniotic cavity of foetal mice between 13 to 16 days post coitus, beta-galactosidase activity was detected between 24 to 120 h after injection. The highest levels of transgene expression were generally observed between 48 to 72 h following injection of the adenoviral vector. We demonstrate that infection of the pulmonary airways is dependent on the developmental stage of the foetus and can be achieved on the 15th day of gestation.
79 citations
TL;DR: Four Lactobacillus strains were tested for their ability to produce and secrete heterologous proteins and it was found that the level of extracellular production was highest in Lb.
Abstract: Four Lactobacillus strains (Lb. plantarum NCIMB 8826, Lb. paracasei LbTGS1.4, Lb. casei ATCC 393 and Lb. fermentum KLD) were tested for their ability to produce and secrete heterologous proteins. These strains were first screened with an alpha-amylase reporter under the control of a set of expression or expression/secretion signals from various lactic acid bacteria. With most of the constructions tested, the level of extracellular production was highest in Lb. plantarum NCIMB 8826, and lowest in Lb. paracasei LbTGS1.4. These two strains were next assayed using a model antigen consisting of the N-terminal part of the M6 protein from Streptococcus pyogenes fused to the linear epitope ELDKWAS from human immunodeficiency virus gp41 protein. Secretion of this heterologous protein was inefficient in Lb. paracasei LbTGS1.4, which accumulated a large intracellular pool of the unprocessed precursor, whereas Lb. plantarum NCIMB 8826 was able to secrete the antigen to a level as high as 10 mg l-1.
64 citations
TL;DR: It is proposed that regulation by TetR-KRAB is a valuable tool with which to study the effects of viral gene expression in vitro and a strong reduction in E2A gene expression, viral DNA replication, and late gene expression was observed in noncomplementing A549 cells.
Abstract: An E1-E3-deleted recombinant adenovirus vector expressing the hybrid protein TetR-KRAB has been produced. In this virus, AdTG9562, the E2 transcription is regulated by TetR-KRAB and tetO sequences inserted in cis. In the absence of tetracycline, a strong reduction in E2A gene expression, viral DNA replication, and late gene expression was observed in noncomplementing A549 cells, and a reduction in viral growth was seen in the E1-expressing 293 cells. In contrast, there was no repression in the presence of the regulator tetracycline. We propose that regulation by TetR-KRAB is a valuable tool with which to study the effects of viral gene expression in vitro.
35 citations
TL;DR: Many important findings in the past year have helped to identify multiple cellular interactions and signals in vertebrates that govern induction of neuroectoderm, its patterning, neural tube formation, and the subsequent differentiation of neurons.
17 citations
Patent•
25 Aug 1997TL;DR: In this article, a method for preparing an homogenous suspension of stable lipid-nucleic acid complexes or particles, comprising of combining one or more cationic lipids, one or many colipids, and stabilizing additives to form a lipid suspension, was presented.
Abstract: The invention is related to a method for preparing an homogenous suspension of stable lipid-nucleic acid complexes or particles, comprising: a) combining one or more cationic lipids, one or more colipids, and one or more stabilizing additives to form a lipid suspension, b) combining the lipid suspension with a nucleic acid to form a complex or a particle, and optionally c) subjecting the complex or particle to a sizing procedure. It also concerns an homogenous suspension produced notably by the above method.
17 citations
Patent•
10 Sep 1997TL;DR: In this article, a method for preparing a plasmid DNA using a wet cell biomass comprising after resuspension of said biomass the following steps:, alkaline lysis, high ionic strength acidification, elimination of solubles, endotoxin and contaminating RNA reduction, filtering gel chromatography and conditioning.
Abstract: The invention concerns a method for preparing a plasmid DNA using a wet cell biomass comprising after resuspension of said biomass the following steps:, alkaline lysis, high ionic strength acidification, elimination of solubles, endotoxin and contaminating RNA reduction, filtering gel chromatography and conditioning. The invention also concerns a pharmaceutical composition containing a plasmid DNA and its use for gene therapy.
5 citations
TL;DR: The results suggest that in vivo transfection mediated by rVV-IL-2 has potential effectiveness in enhancing host immunity and would be a useful approach to cancer gene therapy.
Abstract: Direct gene transfer into somatic tissue iii vivo is a developing technology with potential application for cancer gene therapy. In this study, recombinant vaccinia virus encoding human IL-2 gene (rVV-IL-2) was used as a candidate vector in mediating iii vivo gene therapy. After rVV-IL-2 was expanded in VERO cells for 72 h, high titer (10(8)-10(10) PFU/ml) rVV-IL-2 were harvested. When 10(6) murine melanoma cells (F16-F10) were infected with rVV-IL-2, about 200 U/ml IL-2 activity was detected in the supernatants at 8 h, and the up-regulation of ICAM-1 and MHC-I expressions on the melanoma cells were observed. The treatment of murine melanoma model by local injection of rVV-IL-2 into the tumor site showed that rVV-IL-2 transfection significantly inhibited the tumor growth and prolonged the survival time of tumor-bearing mice. The splenocytes from rVV-IL-2 treated mice showed higher cytotoxicities of NK, LAK and CTL in comparison with those from the controls. These results suggest that in vivo transfection mediated by rVV-IL-2 has potential effectiveness in enhancing host immunity and would be a useful approach to cancer gene therapy.
3 citations
TL;DR: New Phase I clinical trials based on novel protocol design were initiated to improve the safety and efficiency of persistent CFTR gene transfer, and to introduce novel ways of administration and new techniques to assess the potential therapeutic efficacy of functional gene expression.
Abstract: The cloning of the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), mutations of which are responsible for the clinical onset of cystic fibrosis (CF), along with progress in understanding the interplay between CFTR functions and the CF cellular phenotype have prompted many investigators to explore the therapeutic potential of CFTR gene delivery to airway cells in CF patients. In the last four years, a large number of Phase I clinical trials have been started. The results from the very first trials, although mixed, showed that it was possible to transfer and express the CFTR gene, and in certain cases restore the functional electrophysiological properties of the diseased CF cells. These initial trials have been fundamental in encouraging more basic research on vector design to improve the safety and efficiency of persistent CFTR gene transfer, and to introduce novel ways of administration and new techniques to assess the potential therapeutic efficacy of functional gene express...
1 citations
Patent•
25 Mar 1997TL;DR: In this article, a novel antitumour composition including a cell population capable of expressing at least three therapeutic genes, particularly immunostimulatory and/or cytotoxic genes, is disclosed.
Abstract: A novel antitumour composition including a cell population capable of expressing at least three therapeutic genes, particularly immunostimulatory and/or cytotoxic genes, is disclosed. A packaging cell expressing interleukin-2 and a retroviral vector expressing the gamma-interferon and thymidine kinase genes of herpes simplex virus 1 (HSV-1), as well as the therapeutical use thereof for preventing and treating cancer, are also disclosed.