Lymphocyte subset activation

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I am in the process of setting up a flow cytometric method for measuring
lymphocyte subset activation and could use some help.  We have a
FACSCalibur and are interested in using PHA, pokeweed and Con-A as
mitogens; we are also considering using CD2/CD2R as a stimulant.  We are
planning on assessing expression of CD69, CD3, CD4, CD8 and CD19.  Our
main issues involve: 1) stimulation protocols, 2) optimal mitogen
stimulation times and 3) potential differences between adult and
pediatric populations.  If anyone could either share their experiences
and/or suggest a good reference to help us with this process, it would
be much appreciated.

Thank you,

Yvonne G. Dowdy, M.D.

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