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Practical Course HP-F13: Molecular and Immunological Methods in Tumor Virology

Type: Practical Course with Student Seminars

Date: 4.-22. July 2022

Hosts/Supervisors: Martin Müller (responsible organizer contact: martin.mueller@dkfz.de), Marco Binder, Henri-Jacques Delecluse, Susanne Delecluse, Mathias Heikenwälder, Martin Löchelt, Tim Waterboer


Topics and Content:

1st week:
Preparation of virus-like particles from baculovirus infected insect cells and theiranalysis by ultracentrifugation and western blot and ELISA. In addition, AAV gene transfer vectors derived from different serotypes will be titrated using ELISA and PCR. The vectors will be used for gene transfer of reporter genes into cell lines.

2nd week:
Preparation of primary chicken embryo fibroblasts. Infection with influenza virus, haemagglutination assay, plaque assay, immunofluorescence. Bacterial expression of human papillomavirus (HPV) proteins as GST fusions, preparation and characterization of the proteins, use as ELISA antigens to detect human anti-HPV antibodies.

3rd week:
Methods in molecular virology will include vector genome transfection, vector preparation, and transduction of novel target cells. The readout will be marker gene expression and induction of reporter genes in target cells. In parallel, the induction of cellular responses restricting virus replication and vector function will be analyzed and quantified.
To this end a quantitative bioassay for the detection of Interferon (IFN) in the supernatants of virus-infected cells will be performed. This assay is based on the does-dependent protective effect of IFN against lytic infection of a virus-permissive target cell line.

Experiments will be carried out in groups of 2 students. During the course, each student is required to present a seminar with background information relevant to the experiments being performed. Both classical virological techniques as well as modern molecular and cell biological methods will be employed. Scripts and supervision are available in English and in German.

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