Lymphocyte Activation Assays
Howard Shapiro (hms@shapirolab.com)
Thu, 6 Jul 1995 10:25:44 -0400
Bruce Davis cautions Eric Miller about using activation antigen assays to
replace proliferation assays. I would add my caution, although I believe
that some activation antigens may be better than others. CD69, which is
expressed early and transiently, is probably not good for picking up cells
which may already be in early (but not very early) stages of activation,
which would be detected in a proliferation assay. Bromodeoxyuridine should
give the best correlation with thymidine uptake; in studies done many years
ago, my colleagues and I found that the fraction of cells with elevated RNA
content (detectable using 5 micromolar pyronin Y, measuring fluorescence in
the channel normally used for phycoerythrin)correlated well with counts in
proliferation assays. We are about to begin some large-scale studies in
this area, using a multibeam system which will allow us to correlate
phenotype, RNA content, and two or more activation antigens; I think
multiparameter analyses of this type offer the best hope of clarifying the
steps in lymphocyte activation and their clinical significance.
--Howard
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