As I said in a previous message, the CD4+/CD8+ T-cells(CD3+)in our HIV+ patients come in all varieties of brightness. The acquistion rate is in the 100-400 events/second range and I see no evidence of this being a doublet problem in the SSC vs CD45, CD3 vs CD4, or CD3 vs CD8 cytograms. These populations present consistently in patients over time and occasionally are a significant part of the CD+/CD3+ count. Is there any evidence that these dual positive cells (should not be counted as T-helper cells)and does the amount of relative fluorescence of CD4 and CD8 matter? -----Original Message----- From: Jacek Polski [mailto:jpolski@usouthal.edu] Sent: Thursday, November 08, 2001 10:33 AM To: cyto-inbox Subject: Re: RE: cd4 cd8 coexpression This may well be the answer to this intellectually stimulating issue, but the CD4+CD8+ events in my experience (usually below 1%) have the same intensity of CD8 as suppressor cells and slightly lower CD4 expression than helper cells. In my opinion, this observation supports the previously posted notion that CD4 can be expressed on activated CD8+ cells. Regards, Jacek Polski, MD Univ. South Alabama >>> <Alice.L.Givan@dartmouth.edu> 11/06 6:45 PM >>> Hello Flowers, I just wanted to re-inforce Ken Ault's comments about artifactual co-expression of CD4 and CD8 due to coincidence of two cells in the laser beam. Two cells can coincide in the beam either because they are physically aggregated into a clump or because they just happen (by statistical probablility) to be suspended in the same volume of sample buffer as it moves past the laser. A coincidence artifact should be suspected if: 1) as Ken said, the frequency of these CD4/CD8 doubles decreases when the flow rate is decreased (although this may not happen if the cells are in actual clumps). 2) the intensity of each color on the double expressors is the same as the intensity of each color on the relevant single expressors. In other words, if the double expressors form the fourth corner of a perfect rectangle on a dot plot (with the negs, the PE+ singles, and the FITC+ singles forming the other three corners), then you should be suspicious. Alice Alice L. Givan Englert Cell Analysis Laboratory of the Norris Cotton Cancer Center Dartmouth Medical School Lebanon, New Hampshire NH 03756 tel 603-650-7661 fax 603-650-6130 givan@dartmouth.edu
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