Re: CB phenotying

From: Mario Roederer (Roederer@drmr.com)
Date: Fri Mar 03 2000 - 11:55:10 EST


>- In the CD56+ population I also observe 2 intensities: does anyone know
>what these two populations represent? Is it possible that the CD56high
>cells are gamma-delta cells?

No, these are not gamma-delta T cells.

They are both NK populations; the "bright" CD56+ population is
thought to be an early differentiation stage of NK.  The cells are
uniformly CD94+ and do not express other "kir" molecules.

(That's not to say that there aren't CD56+ T cells--but these are
pretty rare and are swamped by the large excess of NK cells in PBMC.
If you gate on CD3+, then you will see the CD56+ T cells.)

>- I see a CD4low population (monocytes) and a reminescent CD4high, but I
>see the same phenomenon with CD8 (sometimes there's a nice segregation
>between the two, but in most cases there isn't): are the CD8low cells NK
>cells?

Some CD8-low cells are NK cells; others are "IEL-like"
(intra-epithelial lymphocyte) T cells that are probably extra-thymic
in origin.  These cells actually express just as much CD8-alpha as
"regular" T cells, but the CD8 is comprised of only CD8a/a homodimers
rather than CD8a/b heterodimers.

The reason they appear less bright in CD8 is that the typical CD8
antibody we use has a much higher affinity for CD8a/b and is titred
with that in mind--so the CD8a/a molecules are not saturated (hence
less bright).  If you put in 10x more CD8 antibody, you will find
that the "CD8-dull" cells are actually just as bright as the
"CD8-bright" cells.

mr



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