Intracellular cytokine staining versus secreted cytokine

From: david.mcfarland@mcmail.vanderbilt.edu
Date: Mon Feb 02 1998 - 18:08:54 EST


Try these:

Jung T, Schauer U, Heusser C, Neumann C, Rieger C:  Detection of intracellular
cytokines by flow cytometry.  J. Immunol. Met. 150:197-207, 1993.

Kabilan L, Anderson G, Lolli F, Ekre H-P, Olsson T, Troye-Blomberg M:  Detection
of intracellular expression and secretion of interferon-gamma at the single-cell
level after activation of human T cells with tetanus toxoid in vitro.  Eur. J.
Immunol. 20:1085-1089,1990.

One problem with intracellular staining seems to be the fact that you can't
distinguish cytokine that is produced by the cell from cytokine that is
internalized by the cell.  In addition, just because a cell is a high producer,
that doesn't necessarily mean it is a high secretor.  Prussin and Metcalfe have
also shown that there is an "extracellular pool" of cytokine that can be
labelled without permeablising the cells.  See:

Prussin C and Metcalfe DD:  Detection of intracytoplasmic cytokine using flow
cytometry and directly conjugated anti-cytokine antibodies.  J. Immunol. Met. 
188:117-128, 1995.

Happy Flowing!


David McFarland
Howard Hughes Medical Institute
Flow Cytometry Facility
Vanderbilt University Medical Center

_______________________________________________________________________________
Subject: Intracellular cytokine staining versus secreted cytokine
From:    Margaret Cooley <m.cooley@unsw.EDU.AU> at +inet
Date:    02/02/1998  2:24 PM


Hi all

Can anyone point me to references where intracellular cytokine staining by
flow cytometry was compared directly to assays (e.g. ELISA) of secreted
cytokine or of cytokine message?  What I am interested in is whether there
is any data correlating frequencies of cytokine producing cells with total
secreted cytokine or cytokine mRNA levels.  I know work relating mRNA
levels to secreted cytokine, but not intracellular cytokine to secreted
cytokine

Thanks for your help.

Margaret A. Cooley
School of Microbiology and Immunology
The University of New South Wales
Sydney, NSW  2052   Australia
Phone:61-2-9385-3761
Fax:  61-2-9385-1591



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