Novel mycobacterial in vitro assay

The objective of this study is - due to the development of a functional in vitro assay - to identify mechanisms that are relevant for control and resistance of Mycobacterium (M.) tuberculosis. At the moment there are no appropriate functional in vitro assays available by which it is possible to investigate the impact of various factors and mechanisms on the host-pathogen-interaction during mycobacterial infections. These processes can now be analysed faster and in more detail due to our novel functional in vitro assay that is based on fluorescent-labeled mycobacteria. The procedure of the new in vitro-assay starts with isolation of monocytes out of PBMCs (Peripheral blood mononuclear cell) done by cell-isolation via MACS. These monocytes are cultured in vitro for a few days to generate so called Monocyte-derived macrophages (MDMs). These MDMs are then infected with a fluorescent-labeled M. bovis BCG reporter strain that contains a Live/Dead (LD) reporter plasmid. After infection the MDMs are co-incubated with activated (T)-cells to analyse their cytotoxic effect on mycobacteria and the infected macrophages.

Researcher: Sven Witkoswki