I am screening products against PBMCs, testing for potential immune activation. I have been looking at CD69 as a good general marker of immune activity, but would prefer something secreted. Does anyone have some good recommendations? Thanks!
You may look for Th1/Th2/Th17 specific cytokines including IL2, IFNg (Th1), IL4, IL5 (Th2) or IL17, IL22 (Th17).
IL2Receptor alpha is a good alternative, and for secreted proteins IL2 is a good candidate, but as Sudeep Kumar points out, many cytokines could be used. But you can also stain for DNA content, the cells will get into cycle, or prior to that they will start enlarging. There is an 8-fold increase in volume of lymphocytes when they get activated..... that ca be seen with some cell counters (I,e. CASY)
Hi Natasha
We use some cell surface markers like CD25, CD38, CD69 and HLA-DR to define activated T-cells. We also use cytokines like IL-2, TNFa, IL4 and IFNg. If you're interested in cytotoxicity as a readout, you could also consider using CD107a, which is a marker of degranulation.
CD71, T9 and transferrin receptor. It is expressed on proliferating cells, reticulocytes, and erythroid precursors. CD71 plays a role in the control of cellular proliferation by facilitating the uptake of iron via ferrotransferrin binding and the recycling of apotransferrin to the cell surface. Is another maker having commercialized CD25, CD38, CD69 At BD in the1980's
Functional characterization of alloreactive T cells identifies CD25 and CD71 as optimal targets for a clinically applicable allodepletion strategy
Sujith Samarasinghe, Christoph Mancao, Martin Pule, Niga Nawroly, Helen Karlsson, Jennifer Brewin, Peter Openshaw, H. Bobby Gaspar, Paul Veys and Persis J. AmroliaBlood 2010 115:396-407; doi: https://doi.org/10.1182/blood-2009-08-235895
"These include negative selection of donor T cells expressing activation markers (eg, CD25, CD69, CD134, CD137, CD147, HLA-DR) "
There is no single secreted maker for immune activation. You should perhaps use some type of multiplex assay, such as one for cytokine profiling. There are many on the market.
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