Any suggestions welcomed on below problem, We are tracking the cytokine profiles (by intracellular staining) produced by CD4+ T cells in patients who have undergone allogeneic bone marrow transplant (BMT). We are looking at cytokine production pre- and post-stimulation of T cells with PHA/ionomycin for 4 hours. We have no difficulty clearly identifying CD4+ T cells on the unstimulated sample, but on the stimulated sample CD4 seems to disappear. CD3 is still clearly evident. Has anyone seen this before? Is the cell fixing process used for ic staining likely to interfere with CD4 expression or anti-CD4 binding? We don't seem to be losing cells in the short culture period. Thanks for any comments David Ritchie
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