RE: ficoll vs lysed blood phenotypes

From: Ronald Rabin (RRABIN@niaid.nih.gov)
Date: Mon Aug 09 1999 - 09:27:57 EST


This reported at a meeting of, I believe, the Clinical Immunology Society.
Ficoll Hypaque decreased B cell percentage to a mean of 35%, but there can
be selective depletion of T and NK populations as well.  The fact that you
see CD5+ B cells may simply reflect the predominance of this type of B cell
in children.

The reference is Brown, M.R., Cleary, J. M.,and Fleisher, T.A.  Selective
lyphocyte depletion by ficoll hypaque density gradient centrifugation.  I am
not sure what meeting this was, or whether this was ever published in a peer
reviewed journal.

								ron


Ronald L. Rabin, M.D.
Clinical Associate
Cytokine Biology Unit, Laboratory of Clinical Investigation
National Institute of Allergy and Infectious Diseases
National Institutes of Health
Bldg. 10/Rm. 11N228
10 Center Drive MSC 1888
Bethesda, MD   20892-1888
Phone:  (301) 402-4910
FAX:      (301) 402-0627


> ----------
> From: 	Te Chih Liu
> Sent: 	Monday, August 2, 1999 12:37 PM
> To: 	Cytometry Mailing List
> Subject: 	ficoll vs lysed blood phenotypes
> 
> 
> This is a question to the clinical users doing immuno-phenotyping in the 
> group.
> 
> Has anyone got a comment to make on the differing phenotypes of 
> mononuclear cells that are sometimes seen when the same sample is 
> prepared using a lysed whole blood approach as opposed to PBMC obtained 
> after a ficoll separation.
> 
> What about marrow samples?
> 
> I'm presently looking at a sample from a 6-year old child with CD10+ ALL. 
> On the ficoll preparation, the plots are that of a straight forward 
> common ALL. On the LWB preparation, there is a distinct CD5+/CD20+ 
> population (15% of MNC), separate from the CD5-/CD20+ B-cells/blasts and 
> the residual CD5+/CD20- T-cells. I can't think of any reason why that 
> population should be there at all.
> 
> Thanks.
> 
> Te-Chih Liu
> 
> 
> 
> Te-Chih Liu, MRCP(UK), MRCPath
> Haematology Division
> National University Hospital
> Singapore
> 
> Phone: (65)-7725353
> Fax: (65)-7751757
> 
> Email: liutc@pacific.net.sg
> 



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