Re: RE: cd4 cd8 coexpression

From: Jacek Polski (jpolski@usouthal.edu)
Date: Thu Nov 08 2001 - 10:32:46 EST


This may well be the answer to this intellectually stimulating issue, but the
CD4+CD8+ events in my experience (usually below 1%) have the same intensity of CD8 as
suppressor cells and slightly lower CD4 expression than helper cells. In my opinion,
this observation supports the previously posted notion that CD4 can be expressed on
activated CD8+ cells.
Regards,
Jacek Polski, MD
Univ. South Alabama


>>> <Alice.L.Givan@dartmouth.edu> 11/06 6:45 PM >>>

Hello Flowers,
I just wanted to re-inforce Ken Ault's comments about artifactual co-expression of CD4
and CD8 due to coincidence of two cells in the laser beam.  Two cells can coincide in
the beam either because they are physically aggregated into a clump or because they
just happen (by statistical probablility)  to be suspended in the same volume of sample
buffer as it moves past the laser.

 A coincidence artifact should be suspected if:
1) as Ken said, the frequency of these CD4/CD8 doubles decreases when the flow rate
is decreased (although this may not happen if the cells are in actual clumps).
2) the intensity of each color on the double expressors  is  the same as the intensity of
each color on the relevant single expressors.  In other words, if the double expressors
form the fourth corner of a perfect rectangle on a dot plot (with the negs, the PE+
singles, and the FITC+ singles forming the other three corners),  then you should
be suspicious.

Alice


Alice L. Givan
Englert Cell Analysis Laboratory
of the Norris Cotton Cancer Center
Dartmouth Medical School
Lebanon, New Hampshire NH 03756
tel 603-650-7661
fax 603-650-6130
givan@dartmouth.edu



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