Search results for "transgene"
showing 10 items of 259 documents
Transgenerationale Effekte belastender Kindheitserlebnisse auf die Borderline-Symptomatik von Jugendlichen
2014
Einleitung: Bisherige Forschungsergebnisse belegen substantielle Zusammenhänge zwischen dem Erleben von Missbrauchs- und Misshandlungserfahrungen in der Kindheit und der Entwicklung der Borderline Persönlichkeitsstörung im Erwachsenenalter. Die Frage nach transgenerationalen Effekten belastender Kindheitserlebnisse sowie möglicher intermittierender Variablen wird hingegen kontrovers diskutiert. Zudem beschäftigten sich bislang wenige Studien mit der Entwicklung von Borderline Symptomen im Jugendalter. An diesen Wissenslücken ansetzend, untersuchen wir die spezifischen Effekte belastender Kindheitserfahrungen von Eltern auf Borderline-Persönlichkeitsmerkmale der Folgegeneration sowie möglich…
PD-1 signalling in CD4+T cells restrains their clonal expansion to an immunogenic stimulus, but is not critically required for peptide-induced tolera…
2010
Summary The ultimate outcome of T-cell recognition of peptide–major histocompatibility complex (MHC) complexes is determined by the molecular context in which antigen presentation is provided. The paradigm is that, after exposure to peptides presented by steady-state dendritic cells (DCs), inhibitory signals dominate, leading to the deletion and/or functional inactivation of antigen-reactive T cells. This has been utilized in a variety of models providing peptide antigen in soluble form in the absence of adjuvant. A co-inhibitory molecule of considerable current interest is PD-1. Here we show that there is the opportunity for the PD-1/PD-L1 interaction to function in inhibiting the T-cell r…
Selection of distinct Valpha/beta T-cell receptor families during in vivo and in vitro T-cell maturation.
1999
The experimental conditions influencing the use of Valphabeta TCR families were examined in lymph node (LN) cells from peptide-immunized C57BL/6 and Vbeta8.2 transgenic mice. Expanded proportions of Vbeta5, Vbeta8.2, Vbeta9, Vbeta12 and Vbeta14 positive cells and an association of Vbeta8.2 with Valpha11 was found in freshly harvested 8-day or 34-day immune LN cells. In contrast, peptide-specific T-cell lines generated in vitro from 8-day immune lymph node cells were found to be almost exclusively of the Valpha2/Vbeta12 family. However, T-cell lines originating from Vbeta8.2 transgenic mice did not show preferential Valpha usage. Anti-Vbeta8.2 antibody produced different effects: when added …
A novel plasmid DNA electroporation method allows transfection of murine DC.
2007
Under steady state conditions dendritic cells (DC) exert tolerogenic function, but acquire potent immunogenic function due to strong upregulation of costimulatory molecules and proinflammatory cytokines. In numerous studies the potential of modified DC to induce tolerance or immune reactions towards a distinct antigen has been demonstrated. However, DC are refractory to transfection with plasmid DNA by non-viral methods. In this study we have tested the suitability of a newly developed electroporation device to transfect immature murine bone-marrow derived DC (BM-DC). Transfected BM-DC expressed reporter molecules at considerable extent which renders this method suitable to perform all kind…
Abstract 3015: Precise investigation of cancer stem cells in mouse glioblastoma
2018
Abstract In this study, we employ mouse models to investigate features and roles of cancer stem cells (CSCs) in glioblastoma (GBM). A nestin-TK-GFP transgene is firstly used to label CSCs in a fully penetrant mouse model of GBM (M7: hGFAP-Cre; Nf1fl/+; p53fl/fl; Ptenfl/+). Food-mediated ganciclovir (GCV) delivery kills proliferative transgene positive cells and significantly prolongs the lives of the transgene bearing mice. Isolation and transplantation of the tumor cells indicates the GFP+ cells are more tumorigenic than the GFP- cells. We then generate and characterize a novel transgene (CGD: nestin-CreERT2-H2BeGFP-hDTR) that labels all the neural stem/progenitor cells in the subventricul…
Oncolytic parvovirus H1 induces release of heat-shock protein HSP72 in susceptible human tumor cells but may not affect primary immune cells.
2003
Certain autonomous parvoviruses preferentially replicate in and kill in vitro-transformed cells and may reduce the incidence of spontaneous and implanted tumors in animals. Hence, these viruses and their derivatives are currently under evaluation as antitumor vectors. However, the mechanisms underlying their tumor-suppressing properties are not yet understood. We asked whether the lytic parvovirus H1 may enhance the immunogenicity of infected tumor cells. Out of human melanoma and gastrointestinal tumor cells, we selected the cell line SK29-Mel-1 being very susceptible to H1-induced apoptotic killing. Here, no upregulation of HLA class I and costimulatory molecules could be observed followi…
Aav-based gene therapy approaches for the treatment of canavan disease
2013
Background: The enzyme Aspartoacylase (ASPA) is normally expressed in oligodendrocytes, the myelin-forming cells in the central nervous system (CNS). ASPA gene mutations cause Canavan Disease (CD), a devastating neurological disorder characterized by psychomotor retardation, and spongiform degeneration of central white matter in affected children. The lack of ASPA leads to the enrichment in its substrate N-acetyl aspartate (NAA) which is a biomarker of CD. With no available treatment and a pathology restricted to the CNS CD has been trialled by gene therapy. However, gene replacement approaches using neurotropic recombinant adeno-associated viral (rAAV) vectors have proved unsuccessful. It …
Morphological characterization of baculovirus Autographa californica multiple nucleopolyhedrovirus
2009
The budded form of baculovirus Autographa californica multiple nucleopolyhedrovirus is used widely in biotechnological applications. In this study, we observed the morphology of baculovirus in nanometer scale by atomic force microscopy. Additionally, the correlation between transduction efficiency and virus stock storage time was evaluated. By atomic force microscopy, asymmetrical baculovirus particles with enlarged head regions were detected. Observed virus stocks contained variable-length particles, 256 ± 40 nm, along with disintegrated particles and/or cellular components. Long-term storage of stocks led to virus aggregation and decreased cellular entry and transgene expression in mammal…
Can Hepatoma Cell Lines be Re-differentiated to be Used in Drug Metabolism Studies?
2013
Knowledge of metabolism, enzymes so far involved, and potential enzyme-inhibiting or enzyme-inducing properties of new compounds is a key issue in drug development. Primary cultured hepatocytes, cytochrome P450 (CYP)-engineered cells and hepatoma cell lines are currently being used for this purpose, but only primary cultures can produce a metabolic profile of a drug similar to that found in vivo and can respond to inducers. Because of their limited accessibility, alternatives to replace human hepatocytes are currently being explored, including the immortalisation of hepatocytes by using different strategies (i.e. SV40 T-large antigen, conditionally immortalised hepatocytes, transfection wi…
A GFP-tagged Muscleblind C protein isoform reporter construct
2010
Drosophila muscleblind (mbl), the ortholog of human Muscleblind-like 1 (MBNL1) gene involved in Myotonic Dystrophy (DM), gives raise to protein isoforms MblA to G. The specific functions and subcellular distribution of isoforms are still largely unknown. To overcome the lack of isoform-specific antibodies we generated transgenic flies that express a GFP:MblC fusion protein under the control of the Gal4/UAS system. The reporter fusion protein was able to functionally complement mbl loss of function mutations, demonstrating activity, and accumulated predominantly in adult muscle nuclei. The fluorescent nature of the reporter makes it appropriate for live imaging detection of MblC protein isof…