I have to get some quick preliminary data, within 45 days, whether my drug effects immunosuppression in an in vivo cancer model.
I have no experience in immunology but have been reading about the critical role of dendritic cells (DCs) in initiating a immune response that can cancer cells exploit DC cells to escape from immune control.
Would the following suggestions be feasible:
1. check for spontaneous apoptosis of blood cells in DC's in peripheral blood mononuclear cells uinsg Annexin-V and TUNEL assays immediately after blood collection, following treatment of my drug in a cancer mouse model.
2. Extract the in vivo tumor and check by IHC the surface markers for DC maturation: CD54, CD80, CD83, CD86 and HLA-DR.
3. in vitro model: treat explant tumor with tumor conditioned medium (TCM) and the drug conditioned medium (DCM) and check by flow cytometry the surface markers for DC maturation.
The above ideas are based on what I have been reading about immunosuppression. I don't have the experimental skills to test suggestion 1 and 3. Is suggestion a good option? Also, can qPCR be done to check any of the DC for surface markers to confirm the effect of my drug on immune response?
Any advice would be much appreciated.