RE: apoptosis in fresh PBMCs

> -----Original Message-----
> From: Marjan [mailto:gharagozlo@pearl.sums.ac.ir]
> Sent: Friday, December 10, 2004 1:08 PM
> To: Cytometry Mailing List
> Subject: apoptosis in fresh PBMCs
> 

> Have you ever examined apoptosis in fresh PBMCs? I read in some 
> articles that apoptosis in fresh PBMCs can not be detected by Tunel 
> test; the cells should be cultured for 3-5 days before apoptosis test.
> What is your idea? I would be grateful if you could kindly 
> give me the 
> benefits of your experiences.

	You will see greater leverls of cell death following a culture
period but it is possible to detect apoptotic cells in fresh PBMCs,
however..  The in vivo mechanisms for clearing apoptotic cells are very
efficient so they are difficult to detect.  Also, the kinetics of the
process demand that the cells express that characteristic that you have
chosen to measure as you r readout. So, for example, the cells may be
triggered to begin the apoptotic cascade in vivo, but not immediately
express certain hallmark features of apoptosis. 		

This reference may help you :  

Niehues T, Mc Closkey TW, Ndagijimana J, Horneff G, Wahn V, and Pahwa S,
Apoptosis in T lymphocyte subsets in HIV infected children measured
immediately ex vivo and following in vitro activation, Clin Diagn Lab
Immunol, 8: 74-78, 2001. 

good luck,
Tom

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Thomas W. Mc Closkey, Ph. D.
Scientist in Charge, Flow Cytometry Facility, North Shore-LIJ Research
Institute
Assistant Professor, Pediatrics, New York University School of Medicine
Biomedical Research Center, 350 Community Drive, Manhasset, Long Island, New
York 11030
ph:  516-562-4844 [office], 516-562-1135/4641 [lab]  fax:  516-562-2866
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