This is generally true. Its hard to get intracellular cytokine staining with mouse cells if it is the first restimultion. It is easier to see cytokine responses to antigen using cloned T cells or an antigen specific line. This is why you see so many PMA stimulations of primed T cells in the mouse literature. I don't have an explanation. --On Thursday, May 25, 2000, 6:32 PM -0500 Helen Horton <hhorton@yakui.primate.wisc.edu> wrote:r > > Dear flowers- > > I am somewhat perplexed. I routinely perform intracellular cytokine > staining on rhesus PBMC where we stimulate with specific antigen (SIV > peptides in our case) to look for Ag-specific T cells responses (both > CD4 and CD8). The protocol works extremely well. A colleague asked me > to perform intracellular staining on mouse splenocytes using the two > peptides which she immunized her mice with. Try as I might I cannot > get antigen-specific IFN-gamma production using my usual protocol > (although with mouse-specific Abs instead of rhesus-specific). I > searched the mailing list archives but could only find examples where > people stimulate with PMA/Ionomycin not with specific antigen. Can > anyone tell me: > > 1) if anyone else has tried antigen-specific intracellular staining > for cytokines in the mouse; and > > 2) why people just use PMA and not specific antigen in their mouse assays > > These may be really dumb questions- I'm just not used to working with > anything except primates! > > Any advice would be greatly appreciated, > > Sincerely, > > Helen > Helen Horton PhD > Wisconsin Regional Primate Research Center > 1220 Capitol Court > Madison > WI 53715 > Tel: (608) 265 3381 > Fax: (608) 263 4031 > hhorton@primate.wisc.edu >
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