(human) CD8 analysis questions

From: bunny (bunny@cotleur.com)
Date: Tue Jun 26 2001 - 10:51:34 EST


friends-

I have a dilemma. I'm participating in a study analyzing
chemokine receptors in MS patients in clinical trials. The
combinations used are:   CD4+/CD45RA+/CHEMOKINE and
CD8+/CD45RA+/CHEMOKINE.

Here is my question: for measuring the CD8/45RA portion, do
I set the markers on background (isotype or
unstained,whatever- that's not my current question) and
accept ALL CD8+ (DIM AND BRIGHT), or specifically CD8 bright+/CD45RA+.

For most of the chemokines, the results are the same (for
either /CD8+ bright/45RA+ or CD8+ dim/45RA+). But for one
marker the results are dramatically different.

{I do understand that the CD8 dims's usually contain NK
cells. I searched both cytometry archives and pubmed, and
found many valuable discussions of the roles of these
various subpopulations. For this study, adding more/or
changing CD markers is out of the question.}

So my main questions here are:
 1)With the combinations described above, WHERE do people
normally set the marker for CD8+ cells;
  and
2) do you then report the data as "%chemokine receptor
expression in activated CD8+45RA+ cells"  ?   or CD8+bright/CD45RA+

--
Bunny Cotleur
Cleveland Clinic Foundation
Neurosciences   NC30
9500 Euclid Avenue

Cleveland, OH  44195

(216) 444-1164
cotleua@ccf.org



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