Jan, My inclination would be say #3. We have just recently come upon what I would describe as a fairly unusual situation in which we see CD16 expression on CD8lo cells in macaque PBMCs (from virally infected animals, by coincidence). These cells have all the classical "signs" of activated cells - increased expression of CD44 & CD95, increased cell size, and granzyme B+. Our original decision to go with CD16 was based on the description that it was expressed on monocytes and not T lymphocytes - but now that appears not to be the case (at least with macaques). Has anyone else seen CD16 expression on T lymphocytes? -Doug Reed -----Original Message----- From: Muellerberghaus, Jan [mailto:muellerberghausj@MSX.UPMC.EDU] Sent: Monday, March 12, 2001 5:47 PM To: cyto-inbox Subject: NK, NK-T, T cell or what? Dear flowers: I am cloning human T cells specific for viral antigens and have slightly confusing result now when I phenotype the clones. They are all CD3+, all CD8+, all CD4-, all HLA-DR+ and now comes the confusing part 10-30% positive for CD56 and 5-10% CD16 positive (the last stimulation with irrradiated feeders and PHA is 12 days ago, the cells are cultured in IL-2 at 1000 U/ml). What are these cells: 1) remaining feeders? 2) my cultures are not clonal? 3) in vitro phenomenon because of culture condition and stimulation? 4) NKT cells? Help with resolving this is appreciated Cheers Jan
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