Re: Opinions on proliferation by flow

From: Ress, S, Stan, Dr (sress@uctgsh1.uct.ac.za)
Date: Tue Aug 17 1999 - 13:20:16 EST


Hi Tony,

One way which works well is to perform a proliferative index 
(PI=S+G2/M) using cell cycle analysis. Cells are permeabilized, fixed 
and stained with propidium iodide. For simultaneous surface analysis, 
FITC-conjugated ab are added before permeabolization.

Take a look at our paper in Clin Exp Immunol 1996, 104: 412-418 (PT 
Lukey et al). The Kinetics of DNA histogram (cell cycle analysis) of 
CD4+ T-cells obtained following antigen stimulation correlated well 
with results obtained using conventional thymidine uptake.

Good luck.

Stan

.....................................................................
 I'd like opinions on the use of flow for the detection of T cell
proliferation. I know that bromodeoxyuridine is an established method,
but right now out of my reach ($$). My thought is to use anti-CD3,
-CD4 and/or -CD8 on bulk lymph node cultures stimulated with or
without antigen to get changes in total numbers of cells.

Thanks,

Tony

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