RE: memory/naive CD8 and CD4 T cells

Cilio:

I refer you to a previous post from Mario Roederer discussing the merits (or
lack of) of using this combination for determining memory vs. naïve T cells.


http://www.cyto.purdue.edu/hmarchiv/2000/2385.htm

 

 

Joanne Lannigan, MS

Director, Flow Cytometry Core Facility

University of Virginia

Jordan Hall, Room 7067

P.O. Box 800734

Charlottesville, VA 22908-0734

Office: 434-924-0274

Lab: 434-243-2695

Fax: 434-982-1071

email: joannelannigan@virginia.edu

-----Original Message-----
From: Corrado Cilio [mailto:Corrado.Cilio@endo.mas.lu.se] 
Sent: Saturday, March 05, 2005 10:55 AM
To: cyto-inbox
Subject: memory/naive CD8 and CD4 T cells

 

Hi all,

I have a discussion going on in the lab concerning the analysis of
memory/naive T cells by flow. We stain human peripheral lymphocytes with
aCD3-FITC, aCD45RA-PE, aCD4- or aCD8-PerC and aCD45RO-APC in whole blood to
determine the memory/naive pheotypes. How people do the analysis? We have
three alternatives on the discussion table and would like to know your
opinion, more to think? 

A. 1st gate: lymphogate SSC,FSC; 2nd gate CD3+CD4+ or CD8+; 3rd dotplot
CD45RA vs CD45RO with quadrant to get the % of different populations. If so
how do you treat the double positives? B. 1st and 2nd gate as above and then
two separate dotpot CD4+ or CD8+ vs CD45RA and CD4+ or CD8+ vs CD45RO to get
the % of all CD45RO+ on CD8+ and CD4+. C. 1st and 2nd gate as above and then
using histogram instead of the last dotplots.

Thank you for some suggestions and opinions, hoping to add some more spice
to the discussion.

Corrado

Corrado M. Cilio, M.D., Ph.D. 
Assistant Professor 
Cellular Autoimmunity Unit 
Dept. of Clinical Sciences/Paediatrics 
Malmo University Hospital 
Lund University 
205 02 Malmo, Sweden 
Tel: office +46-40332395; mobil: +46-70-4330338 
"Judge a men by his questions rather than his answers" - Voltaire