I have been using the CD25/OX40 AIM assay for identification of SIV-specific CD4 T cells in rhesus macaques in T cells stimulated with 1mg/ml of Env pooled peptide. I used this concentration to stimulate LN CD4 T cells for 40 hours and OX40 presented good detection efficacy in LN samples. When I performed the same assay in PBMCs with exact Env peptide concentration and stimulation time, CD25 responded well but OX40 did not. In this case:
* can I use only CD25 to define my Env-specific population?
* what might be causing OX40 limitations in detecting Env specific T cells in blood of rhesus macaques?
* Is there any marker that would work better than Ox40 in rare circulating Ag specific T cell populations?
Many thanks in advance for the help.
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