I can't help but interject one caution to generally applying this. Gamma delta T-cells can be CD4-CD8- and can be a significant population in Asian patients and setting of mycobacterial infection. In addition, there are disease processes where there are high numbers of alpha beta T-cells that are CD4-CD8-- eg ALPS. Sooooo- this is a useful approach to detect recently CD4 positive T-cells in specific situations but is not generally applicable. Maryalice >Kathy, > >Dr. O'Gorman in Chicago has a nice technique for accomplishing this. > >He stains with CD3 and CD8, then gates the CD3+, CD8- events and assumes these >to be the cells which would express CD4 (has they not been overstimulated). > >I believe he described this in a paper concerning a CD40L assay he developed. > >Keith Bahjat >Graduate Assistant >University of Florida >kbahjat@ufl.edu > > >Kathleen Schell wrote: > >> Dear Flowers, >> We have an investigator who is studying cytokine production in >> response to mitogenic stimulus. We observe a down regulation of the CD4 >> molecule ( in surface staining) by 24 hours that renders this population >> unidentifiable. They are looking at intracellular cytokines, and surface >> molecules are stained first, cells are fixed, permeabilzed, and then >> stained for cytokines. Has anyone seen this? We have repeatedly seen a >> decrease in the fluorescence of CD3 after PHA stimulation, but never to the >> point where the population can no longer be seen--this has been reported in >> the literature. Is there another way to identify the CD4 population using >> this kind of staining protocol? Cells not stimulated have good surface >> expression of CD4. >> Thanks in advance. >> Kathy >> >> -------------------------------------------------------------- >> Kathy Schell >> Supervisor, UWCCC Flow Cytometry Facility >> 600 Highland Ave. K4/535 >> Madison, WI 53792 >> Voice: 608-263-0313 >> e-mail: kschell@facstaff.wisc.edu Maryalice Stetler-Stevenson Director Flow Cytometry Unit Laboratory of Pathology, NCI, NIH
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