Practical Course HP-F13: Molecular and Immunological Methods in Tumor Virology
Type: Practical Course with Student Seminars
Date: 8.-26. July 2024
Hosts/Supervisors: Martin Müller (responsible organizer contact: martin.mueller@dkfz.de), Stella Autenrieth, Marco Binder, Timo Bund, Henri-Jacques Delecluse, Susanne Delecluse, Aoife Gahlawat, Tim Waterboer
Topics and Content:
General
Order of weeks is variable. Experiments will be carried out in groups of 2-3 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.
Week 1 (M. Müller & T. Waterboer/A. Gahlawat):
Preparation of virus-like particles from baculovirus infected insect cells and their analysis by ultracentrifugation and Western blot and ELISA.
Processing of cancer patient blood and biobanking. Multiplex serology for the analysis of cytokines in HPV+ cancer.
Week 2 (S. Autenrieth & M. Binder):
Innate immune responses to viral infection will be investigated. Cell-intrinsic induction of antiviral defenses will be studied in virus infected A549 epithelial cells and their supernatants. Furthermore, immune cells (PBMCs) will be stimulated by incubation with bacteria or a virus, or by co-cultivation with virus infected A549 cells. Supernatants will be analyzed for presence of cytokines by ELISA, and for their antiviral activity in a bioassay. Lastly, activation of immune cells will be analyzed by spectral flow cytometry (an advanced version of FACS).
Week 3 (H.-J & S. Delecluse; T. Bund):
The first practical (days 1+2) will use life cell imaging to characterize cells infected with the Epstein-Barr virus.
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.