Flow Cytometer/Second Mice Experiment
Immunity Project is still in the process of setting up our Flow Cytometer. It’s in a lot of pieces – our brain trust is putting it together.
We have exciting news to report. We were able to negotiate significant additional discounts with our vendors, so we are now adding an additional experiment to further validate our approach. The new experiment is essentially the same as the first but with blood from a donor with a different HLA type. This will allow us to look at and vaccinate the (original) HLA(B57) mice with the KF11 epitope and vaccinate the (new) HLA(B27) mice with the KK10 epitope.
We will be looking for an immune response using an interferon gamma elispot assay. To do this, we will take splenic blood from the mice and incubate with the epitope used for vaccination and count the spot forming cells (SFCs) per million peripheral blood monocytes. Currently we run the elispot assay by setting up the plates in our lab and sending them out for quantitative counts of the SFCs. We keep high-resolution photographs of the plates for our records.
In addition to the Elispot data, there are a lot of things we can look at with our flow cytometer to follow the various events we expect to see happening like antigen presentation and T-Cell activation. We also got a great suggestion from hacker news to explore MHC staining to follow the antigen presentation process happening in our experiments. We found off-the-shelf staining kits from Proimmune for the KK10 and SL09 epitopes – we are trying to get those reagents into the lab now.