RE[2]: markers to distinguish mouse monocytes

From: Weaver, James L (WEAVER@CDER.FDA.GOV)
Date: Wed Sep 18 2002 - 14:56:18 EST


I agree with Julie that CD11b and Gr-1 are the best markers currently
available to resolve the phagocytic populations in mouse peripheral blood.
The situation may be more complex. Attached is a contour plot of 250,000
CD45-PE/Cy5 bright leukocytes stained with Gr-1-FITC and CD11b-PE  using a
no-lyse/no wash protocol (1). Depending on ones degree of bravery, there are
5-7 populations that can be identified. Populations A/B (Gr-1 & CD11b
bright) are granulocytes, I have noted that the proportion in B is larger
after erythrocyte lysis. Pops C & D (Gr-1 neg, CD11b medium & high) are
monocytes and based on (2), likely a subpopulation of B-cells although I do
not know which ones are which. Pop E (Gr-1 low, CD11b neg) is probably a
T-cell subpopulation (2). Population F (Gr-1 & CD11b low) is unknown but the
peak increases as cells with lower CD45 expression are included in the gate.
Pop G, (CD11b & Gr-1 neg) is the bulk of T & B lymphocytes in peripheral
blood.

-Jim Weaver

Refs
(1) Serial Phenotypic Analysis of Mouse Peripheral Blood Leukocytes. James
L. Weaver, Dennis D.  Broud, Katherine McKinnon, and Dori R. Germolec.
Toxicol. Mech. Methods 12 95-118 (2002).

(2) Flow cytometric identification of murine neutrophils and monocytes. Eric
Lagasse & Irving Weissman. J. Immunol. Meth. 197 139-150 (1996).

*  -----Original Message-----
*  From: Oughton, Julie [mailto:julie.oughton@orst.edu]
*  Sent: Wednesday, September 11, 2002 6:44 PM
*  To: Cytometry Mailing List
*  Subject: RE: markers to distinguish mouse monocytes
*
*
*
*  Oops!
*
*  I misspoke in my message re: the best markers to identify
*  monocytes in mouse
*  blood. I still stand by CD11b and Gr-1 as the best markers to resolve
*  monocytes from granulocytes. As you will see from the
*  histograms illustrated
*  in the attached word document, both monocytes and
*  granulocytes express CD11b
*  and Gr-1. However, granulocytes express high levels of Gr-1
*  while monocytes
*  express low levels of Gr-1 [I had previously said that only
*  granulocytes
*  express Gr-1, my mistake]. I used color eventing to show the
*  profiles of
*  these two populations.
*
*  Sorry for the mistake!
*
*  Julie Oughton
*  Oregon State University
*
*
*
*   <<CD11b Gr-1.doc>>
*



Mouse_Blood_Phagocytes.jpg



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