JoVE-Enzyklopädie der Experimente
Mikrobiologie
0 Aufrufe • 3:02 Min. • July 1st, 2026
Begin with a single-cell suspension of splenocytes, harvested from a Listeria monocytogenes-infected mouse.
The suspension contains immune cells, including antigen-presenting cells and T cells.
The cells are suspended in a culture medium containing serum proteins to support cell viability.
Now, seed the suspension into a multi-well plate containing heat-killed L. monocytogenes.
During incubation, antigen-presenting cells engulf and process the bacteria, then display the bacteria-derived antigens on MHC molecules.
T-cell receptors recognize and bind to these bacterial antigens on MHC,
This activates the T cells and triggers cytokine release.
Next, add a protein inhibitor of vesicular transport and incubate.
Inside T cells, the inhibitor blocks vesicular transport, preventing further cytokine release.
Collect the culture medium containing cytokines.
Add a buffer and collect the cells separately.
The cytokine-containing culture medium and harvested cells are ready for downstream analysis to assess the L. monocytogenes-induced cytokine production in T cells.
To measure CD4 and CD8 T cell interferon gamma responses, harvest the spleens from mice infected with two times ten to the fourth CFU of the
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