I've got a question for the list regarding CD69 expression, or rather the inability on our part to detect it. We've been staining primate (cyno) PBMC, looking at the host response to viral infection. Everything looks great, but nowhere do we see any CD69 expression. Compared to negative controls we see a slight - very slight, by my eyes - shift, nothing I'd want to write home about. Back a few years ago when I was knee-deep in mice in Connecticut we tried it with mice and also saw very poor staining - again, hardly anything I was impressed with at the time. I look in the literature, however, and I see beautiful CD69 staining. I've left this alone for a while, not really a front-burner of an issue, but it has lurked there out of sight, waiting for me to pick it up again. Last night I sat down and read a paper in JI from Louis Picker's group in Oregon, looking at memory cells in the rhesus macaque. Looking at the methods it clearly states that for intracellular cytokine staining the cells are stained for surface antigens, fixed & permeablized, and then stained with CD69 and cytokine mAbs. So I'm wondering - is that where I've been going wrong? Any thoughts? I tried searching the archives but most of the messages pulled up by CD69 deal with it's relationship with proliferation. Thanks! Doug Douglas S. Reed, Ph.D. Microbiologist Respiratory & Mucosal Immunity Department of Aerobiology & Product Evaluation Division of Toxinology & Aerobiology U.S. Army Medical Research Institute of Infectious Diseases 1425 Porter Street, Fort Detrick Frederick, MD 21702-5011 301-619-6728 301-619-6911 fax doug.reed@det.amedd.army.mil
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