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The Immune System – Multimedia

Biosynthesis of Very Long Chain Polyunsaturated Fatty Acids in Chicory content piece image
Poster

Biosynthesis of Very Long Chain Polyunsaturated Fatty Acids in Chicory

Very long chain polyunsaturated fatty acids (VLCPUFAs) are declining in reserves, since they are produced by oily fish. VLCPUFAs are important in human nutrition because they are responsible for the production of inflammatory mediators such as prostaglandins, the regulation of cholesterol synthesis and its transport, and the maintenance of cellular membranes. This research aims to produce VLCPUFAs in edible plants.
Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer content piece image
Poster

Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer

Constitutive TNF-alpha expression is characteristic of the malignant ovarian surface epithelium. Adenoviral mutants hold great promise as gene therapy vectors but their efficacy is hindered by an inflammatory cascade orchestrated by TNF-alpha. We found that delivering TNF-alpha shRNA to ovarian cancer cells using oncolytic adenoviruses could reduce the inflammatory cascade generated by adenoviruses and also had direct anti-tumour activity on the cancer cells.
Highly Efficient High-Throughput Transfection content piece image
Poster

Highly Efficient High-Throughput Transfection

Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines content piece image
Poster

Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines

The amaxa Nucleofector® Technology is a well established method for effective, non-viral transfection of any nucleic acid substrate into hard-to-transfect cells, especially suspension and primary cells. With the Nucleofector® 96-well Shuttle® system high throughput applications such as siRNA-library screenings have become amenable for the first time in these cell types. This renders target validation and identification possible in cell types highly relevant for biomedical research. Here we dis
Development of an Automated siRNA Screening of Host Macrophages Genes Involved in Mycobacterium Tuberculosis Infection content piece image
Poster

Development of an Automated siRNA Screening of Host Macrophages Genes Involved in Mycobacterium Tuberculosis Infection

In order to identify host genes required for M. tuberculosis infection and persistence, we developed a phenotypic cell-based assay in both murine and human cells and adapted it for high throughput and high content screening purposes. Knock-down efficiencies above 80% were achieved in “hard-to-Transfect” macrophage cells. Validation of the assay performed with control siRNAs will be discussed.
Antibody Array-Based Analysis of Expression Levels in Protein Mixtures Extracted from Formalin-Fixed Paraffin-Embedded (FFPE) Material using ULS Labeling content piece image
Poster

Antibody Array-Based Analysis of Expression Levels in Protein Mixtures Extracted from Formalin-Fixed Paraffin-Embedded (FFPE) Material using ULS Labeling

Protein was extracted from 7-year old FFPE samples of B-cell lymphoma and mamma carcinoma, followed by ULS labeling. Samples were used for single or two color assays using a custom-made antibody array. We observed differential expression levels for several captured analytes. In B-cell lymphoma, expected high levels of IgM were detected, while high levels of CA19-9, a putative marker, were found for the breast cancer tissue.
Determination of the Autoantibody Repertoire in Autoimmune Diseases with Factor Protein Arrays – A new Approach to Personalized Therapies content piece image
Poster

Determination of the Autoantibody Repertoire in Autoimmune Diseases with Factor Protein Arrays – A new Approach to Personalized Therapies

Recently, we have presented a strategy to identify new marker molecules characteristic for Alopecia areata by combining protein microarray technology with the use of large cDNA expression libraries (Lueking et al., 2005). This strategy is now applied to Juvenile idiopathic Arthritis (JIA), Rheumatoid Arthritis (RA) and Multiple Sclerosis (MS) in a BMBF-funded project.
Fluorometric Assays for the Evaluation of Chemokine Receptor Inhibitors with Anti-HIV Activity content piece image
Poster

Fluorometric Assays for the Evaluation of Chemokine Receptor Inhibitors with Anti-HIV Activity

In order to infect a target cell, the HIV envelope glycoprotein gp120 has to interact with both the cellular CD4 receptor and a chemokine receptor, CCR5 or CXCR4, the so-called HIV co-receptors. Several flow cytometric/fluorometric assays are developed in our lab and are very helpful in deciphering the mode of action and interaction site of several classes of HIV entry inhibitors, such as the chemokine receptor antagonists.
LacZ Reporter Fusion Assay for Rapid and Easy Identification of Highly Efficient siRNA and its Delivery by Lentivirus into Suspension Cell Lines content piece image
Poster

LacZ Reporter Fusion Assay for Rapid and Easy Identification of Highly Efficient siRNA and its Delivery by Lentivirus into Suspension Cell Lines

The LacZ reporter fusion assay is an excellent method to precisely quantify knockdown efficiency of siRNA-sequences and can be corroborated by Western blot analysis. Efficient production of Lentiviruses and high transduction efficiency (up to 98%) were achieved on lymphoid B- and T-cells as demonstrated by FACS analysis and GFP expression. Successful knockdown effect with specific shRNA was observed in suspension cells three days after lentiviral infection.
Quantitative Evaluation of High-Abundance Serum Proteins Depletion  content piece image
Poster

Quantitative Evaluation of High-Abundance Serum Proteins Depletion

Identification of low abundant proteins requires relative enrichment. We have tested systems for specific removal of high abundance proteins. Efficiency of albumin and IgG removal was assessed by ELISA and 2-DE. To estimate unspecific binding, six cytokines were spiked into serum. Recovery rates were determined with a cytometric bead assay.
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