Immunosuppression - (Jan/20/2015 )

Hi,

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.

I think all your ideas are good and testable . If the surface markers are modulated at gene expression level, you can use qPCR. 

But I am sure you do realize that all these experiments need thorough plans with great considerations for positive and negative controls. Best place to start would be to read some relatively recent papers who have done similar work, and not rush it because you have 45 days.

Thanks. I'm just wondering that if the DC cells are so important in immune response why are there no reports on the serum levels of markers of DC maturation: CD54, CD80, CD86 etc.

Also, what are other simple methods to observe change in immune response? I was thinking of serum IL-6, IL-7, IL8, sHLA-G etc

These are good questions: But I think you are looking at a local (tumor) phenomenon and hoping to see a system-level change (serum levels), so,  if I have to guess, I would say, 

1. Changes are likely not detectable

2. Detectable changes are likely non-specific