<ol>
	<li>Khurana, A., Kronenberg, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Method+for+Production+of+Recombinant+CD1d+Protein+in+Insect+Cells.">A Method for Production of Recombinant CD1d Protein in Insect Cells.</a> 12/10/2007) Journal of Visualized Experiments. , .</li></ol>

The authors have nothing to disclose.

interview: glycolipid antigen presentation by cd1d and the therapeutic potential of nkt cell activation

Natural Killer T-Zellen (NKT) sind kritische Faktoren für die Immunantwort gegen Krebs, die Regulierung des autioimmune Krankheit, Clearance von Infektionserregern und die Entwicklung von artheriosklerotischer Plaques. In diesem Interview spricht Mitch Kronenberg seinem Labor die Bemühungen um den Mechanismus, durch die NKT-Zellen durch Glycolipid Antigene aktiviert werden, zu verstehen. Im Mittelpunkt dieser Studien ist CD1d - die Antigen-präsentierende Molekül, Glykolipide präsentiert NKT-Zellen. Das Aufkommen der CD1d Tetramer-Technologie, eine Technik, durch die Kronenberg Labor entwickelt, ist entscheidend für die Sortierung und Kennzeichnung von Teilmengen von spezifischen Glycolipid-reaktiven T-Zellen. Mitch erklärt, wie Glycolipid-Agonisten wird als therapeutische Wirkstoffe verwendet werden, um NKT-Zellen bei Krebspatienten zu aktivieren und wie CD1d Tetramere können verwendet werden, um den Zustand der NKT-Zell-Population in vivo folgenden Glycolipid-Agonisten-Therapie zu beurteilen. Aktueller Stand der laufenden klinischen Studien mit diesen Agonisten als auch Mitch Vorhersage für Bereiche, in dem Gebiet der Immunologie, die haben zunehmende Bedeutung in der nahen Zukunft diskutiert.

Watch this Scientific Journal Video about Interview: Glycolipid Antigen Presentation by CD1d and the Therapeutic Potential of NKT cell Activation at JoVE.com

Interview: Glycolipid Antigen Presentation by CD1d and the Therapeutic Potential of NKT cell Activation

Natural Killer T-Zellen (NKT) sind kritische Faktoren für die Immunantwort gegen Krebs, die Regulierung des autioimmunity, Clearance von Infektion und der Entwicklung von artheriosklerotischer Plaques. In diesem Interview spricht Mitch Kronenberg seinem Labor die Bemühungen um den Mechanismus, durch die NKT-Zellen durch Glycolipid Antigene aktiviert werden, zu verstehen.

Interview: Glycolipid Antigen-Präsentation durch CD1d und das therapeutische Potenzial von NKT Zellen-Aktivierung

Natural Killer T cells (NKT) are critical determinants of the immune response to cancer, regulation of autioimmune disease, clearance of infectious ...

interview-glycolipid-antigen-presentation-cd1d-therapeutic-potential

Research

JoVE Journal

Biology

9.8K Aufrufe.  La Jolla Institute for Allergy and Immunology. Natural Killer T-Zellen (NKT) sind kritische Faktoren für die Immunantwort gegen Krebs, die Regulierung des autioimmunity, Clearance von Infektion und der Entwicklung von artheriosklerotischer Plaques. In diesem Interview spricht Mitch Kronenberg seinem Labor die Bemühungen um den Mechanismus, durch die NKT-Zellen durch Glycolipid Antigene aktiviert werden, zu verstehen.

Serie: Interview: Glycolipid Antigen Presentation by CD1d and the Therapeutic Potential of NKT cell Activation

Christopher Hughes:  An in vitro model for the Study of Angiogenesis (Interview)

Christopher C.W. Hughes describes the utility of his culture system for studying angiogenesis in vitro.  He explains the importance of fibroblasts that secrete a critical, yet unidentified, soluble factor that allow endothelial cells to form vessels in culture that branch, form proper lumens, and undergo anastamosis.

Christopher Hughes:  An in vitro model for the Study of Angiogenesis Interview

Christopher C.W. Hughes describes the utility of his culture system for studying angiogenesis in vitro. He explains the importance of fibroblasts that secrete a critical, yet unidentified, soluble factor that allow endothelial cells to form vessels in culture that branch, form proper lumens, and undergo anastamosis.

Christopher Hughes: Eine In-vitro-Modell für das Studium der Angiogenese (Interview)

Christopher C.W. Hughes describes the utility of his culture system for studying angiogenesis in vitro.  He explains the importance of fibroblasts that ...

Forschung

Biologie

Investigating the Microbial Community in the Termite Hindgut - Interview

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes.     The symbiotic relationship existing between the host insect and lignocellulose-degrading gut microbes is explained, as well as the industrial uses of these microbes for degrading plant biomass and generating biofuels.

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes. The symbiotic relationship existing between the host insect and lignocellulose-degrading gut microbes is explained, as well as the industrial uses of these microbes for degrading plant biomass and generating biofuels.

Untersuchung der mikrobiellen Gemeinschaft in der Termite Enddarm - Interview

Jared Leadbetter explains why the termite-gut microbial community is an excellent system for studying the complex interactions between microbes.     The ...

Microbial Communities in Nature and Laboratory - Interview

Mikrobielle Gemeinschaften in Natur und Labor - Interview

Studies of Bacterial Chemotaxis Using Microfluidics - Interview

Studium der bakteriellen Chemotaxis Mit Mikrofluidik - Interview

Biology of Microbial Communities  - Interview

Biologie mikrobieller Gemeinschaften - Interview

Regulatory T cells: Therapeutic Potential for Treating Transplant Rejection and Type I Diabetes

Regulatorische T-Zellen: Therapeutische Potenzial zur Behandlung von Transplantatabstoßung und Typ-I-Diabetes

Interview: Protein Folding and Studies of Neurodegenerative Diseases

In this interview, Dr. Lindquist describes relationships between protein folding, prion diseases and neurodegenerative disorders. The problem of the protein folding is at the core of the modern biology. In addition to their traditional biochemical functions, proteins can mediate transfer of biological information and therefore can be considered a genetic material. This recently discovered function of proteins has important implications for studies of human disorders. Dr. Lindquist also describes current experimental approaches to investigate the mechanism of neurodegenerative diseases based on genetic studies in model organisms.

In this interview, Dr. Lindquist describes relationships between protein folding, prion diseases and neurodegenerative disorders. The problem of the protein folding is at the core of the modern biology. In addition to their traditional biochemical functions, proteins can mediate transfer of biological information and therefore can be considered a genetic material. This recently discovered function of proteins has important implications for studies of human disorders. Dr. Lindquist also describes current experimental approaches to investigate the mechanism of neurodegenerative diseases based on genetic studies in model organisms.

Interview: Proteinfaltung und Studien für Neurodegenerative Erkrankungen

In this interview, Dr. Lindquist describes relationships between protein folding, prion diseases and neurodegenerative disorders. The problem of the ...

Interview: Bioreactors and Surfaced-Modified 3D-Scaffolds for Stem Cell Research

A Nature Editorial in 2003 asked the question "Good-bye, flat biology?" What does this question imply? In the past, many in vitro culture systems, mainly monolayer cultures, often suffered from the disadvantage that differentiated primary cells had a relatively short life-span and de-differentiated during culture. As a consequence, most of their organ-specific functions were lost rapidly. Thus, in order to reproduce better conditions for these cells in vitro, modifications and adaptations have been made to conventional monolayer cultures.
The last generation of CellChips -- micro-thermoformed containers -- a specific technology was developed, which offers the additional possibility to modify the whole surface of the 3D formed containers. This allows a surface-patterning on a submicron scale with distinct signalling molecules. Sensors and signal electrodes may be incorporated. Applications range from basic research in cell biology to toxicology and pharmacology. Using biodegradable polymers, clinical applications become a possibility. Furthermore, the last generation of micro-thermoformed chips has been optimized to allow for cheap mass production.

In the past many in vitro culture systems -- mainly monolayer cultures -- often suffered from the disadvantage that differentiated primary cells had a relatively short life-span and de-differentiated during culture. As a consequence, most of their organ-specific functions were lost rapidly. Thus, in order to reproduce better conditions for these cells in vitro, modifications and adaptations have been made to conventional monolayer cultures.

Interview: Bioreaktoren und Aufgetaucht-Modified 3D-Gerüste für Stammzellforschung

A Nature Editorial in 2003 asked the question  "Good-bye, flat biology?"  What does this question imply? In the past, many in vitro culture systems,  ...

Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase

HIV-1 integrates into the host chromosome of infected cells and persists as a provirus flanked by long terminal repeats. Current treatment strategies primarily target virus enzymes or virus-cell fusion, suppressing the viral life cycle without eradicating the infection. Since the integrated provirus is not targeted by these approaches, new resistant strains of HIV-1 may emerge. Here, we report that the engineered recombinase Tre (see Molecular evolution of the Tre recombinase , Buchholz, F., Max Planck Institute for Cell Biology and Genetics, Dresden) efficiently excises integrated HIV-1 proviral DNA from the genome of infected cells. We produced loxLTR containing viral pseudotypes and infected HeLa cells to examine whether Tre recombinase can excise the provirus from the genome of HIV-1 infected human cells. A virus particle-releasing cell line was cloned and transfected with a plasmid expressing Tre or with a parental control vector. Recombinase activity and virus production were monitored. All assays demonstrated the efficient deletion of the provirus from infected cells without visible cytotoxic effects. These results serve as proof of principle that it is possible to evolve a recombinase to specifically target an HIV-1 LTR and that this recombinase is capable of excising the HIV-1 provirus from the genome of HIV-1-infected human cells.
Before an engineered recombinase could enter the therapeutic arena, however, significant obstacles need to be overcome. Among the most critical issues, that we face, are an efficient and safe delivery to targeted cells and the absence of side effects.

Current HIV-1 strategies act to suppress the viral life cycle but do not effectively eradicate infection. Here, we demonstrate that an engineered recombinase can efficiently excise integrated HIV-1 proviral DNA from the genome of infected cells.

Interview: HIV-1 Provirus-DNA Excision Mit einem Evolved Rekombinase

HIV-1 integrates into the host chromosome of infected cells and persists as a provirus flanked by long terminal repeats. Current treatment strategies ...

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1. Apoptosis can be triggered when cells encounter a wide range of cytotoxic stresses. These insults initiate signaling cascades that ultimately cause the release of cytochrome c from the mitochondrial intermembrane space to the cytoplasm2. The release of cytochrome c from mitochondria is a key event that triggers the rapid activation of caspases, the key cellular proteases which ultimately execute cell death3-4.
The pathway of apoptosis is regulated at points upstream and downstream of cytochrome c release from mitochondria5. In order to study the post-mitochondrial regulation of caspase activation, many investigators have turned to direct cytoplasmic microinjection of holocytochrome c (heme-attached) protein into cells6-9. Cytochrome c is normally localized to the mitochondria where attachment of a heme group is necessary to enable it to activate apoptosis10-11. Therefore, to directly activate caspases, it is necessary to inject the holocytochrome c protein instead of its cDNA, because while the expression of cytochrome c from cDNA constructs will result in mitochondrial targeting and heme attachment, it will be sequestered from cytosolic caspases. Thus, the direct cytosolic microinjection of purified heme-attached cytochrome c protein is a useful tool to mimic mitochondrial cytochrome c release and apoptosis without the use of toxic insults which cause cellular and mitochondrial damage.
In this article, we describe a method for the microinjection of cytochrome c protein into cells, using mouse embryonic fibroblasts (MEFs) and primary sympathetic neurons as examples. While this protocol focuses on the injection of cytochrome c for investigations of apoptosis, the techniques shown here can also be easily adapted for microinjection of other proteins of interest.

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

In this protocol, we describe the direct cytoplasmic microinjection of cytochrome c protein into fibroblasts and primary sympathetic neurons. This technique allows for the introduction of cytochrome c protein into the cytoplasm of cells and mimics the release of cytochrome c from mitochondria, which occurs during apoptosis.

Die Aktivierung der Apoptose durch cytoplasmatische Mikroinjektion von Cytochrom C</em

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1.  Apoptosis can be triggered when cells encounter a ...