We have a rabbit model system to develop vaccines. Because of the limitation of reagents for rabbit T cell culture, we have to optimize our in vitro culture conditions all the time. Nonspecific T cells are one of the problems in our cultures.
I have not worked with Rabbit CTLs. But in the mouse and swine systems, you normally have to culture your primed CD8+ T cells with APCs presenting the specific antigen for your CTLs. This is done for at least 5 days with or without addition of IL-2. At the end of this period you should end up with antigen-specific CD8+ T cells as majority. You could check this if you had MHC class I tetramers. But for sure you should check the IFNg production in CD8+ T cells (intracellular staining assay and flow cytometry).These cells at this stage produce appreciable levels IFNg.
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