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T helper responses - (Apr/09/2012 )

We did an animal experiment and one of the goals was to look for any changes in T cell activation (primarily Th1 vs Th2 vs Th17). We did a luminex assay to assess for various cytokines on the plasma, but almost all were below the limit of detection, so unfortunately no data was obtained. We saved and froze the spleen and a bunch of lymph nodes. Is anyone aware of any good protocols to look directly at the immune cells themselves to suggest activation of any T helper responses from these tissues? Thank you guys so much in advance.


Did you mean analyze frozen samples? If not, you could take splenocytes and LN cells, stimulate them in vitro with aCD3/CD28 or beads and then do intracellular cytokine staining. That should give you an idea of the various T helper subsets.