Re: Granulocytes fr monocytes by Flow

From: Ronald Rabin (rr84g@nih.gov)
Date: Mon Oct 07 2002 - 17:28:36 EST


Woong-Ki, When I was doing a lot of flow on monocytes and granulocytes, it
was my understanding that unlike lymphoid cells, myelocytic differentiation
markers are not exclusively expressed on one population vs. another.  Thus,
unlike CD19 on B cells vs. CD3 on T cells, CD14 is highly expressed on
monocytes and expressed on lower levels on granulocytes, and the converse is
true for CD15.

That being said, differential levels of expression of these two markers
should allow very good discrimination between the two cells.  In addition,
CD16 is very brightly expressed on all granulocytes, and is at lower levels
on a subset of monocytes.  I think that there are differences between the
isoform of CD16 expressed on these cells, the granulocytes expressing the PI
glycolinked form recognized by 3G8, and monocytes expressing another form.
(I am not sure of this, you will need to check it out).

According to the BD chart, CD80 and CD86 are expressed on monocytes and not
granulocytes.

ron

on 10/4/02 12:39 PM, wkim1@caregroup.harvard.edu at
wkim1@caregroup.harvard.edu wrote:

>
> Dear Flowers,
>
> We are looking for a (or the) marker for granulocytes in human or monkey
> PBMC to distinguish monocytes from them.  Gating monocytes in our Ficolled
> PBMC based upon FSC/SSC plot alone does not do the job (cannot exclude
> contaminating granulocytes, I believe).  The purpose of our PBMC prep is not
> to eliminate granulocytes, but to evaluate granulocytic contribution to
> monocyte population (i.e. enumerating granulocytes vs. monocytes).  To serve
> this purpose, we need to have a pan-marker for human and monkey granulocytes
> if there is such an antibody.  I would very much appreciate for your help
> and expertise.
>
> Sincerely,
>
> Woong-Ki Kim, Ph.D.
> BIDMC
> Div Viral Pathogenes
>

Ronald L. Rabin, M.D.
Senior Staff Fellow
Laboratory of Immunobiochemistry
DBPAP/OVRR
Center for Biologics Evaluation and Research
U.S. Food and Drug Administration
29 Lincoln Drive (MSC-4555)
Building 29, Room 129
Bethesda, MD   20892-4555

phone:  301.496.8806
fax:    301.402.5177
email:  rr84g@nih.gov



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