Hi,
I am at CD8 cell priming following co-culture with stimulated murine DCs.
I wondered if anyone could suggest a cell line for this, or might have any they would be willing to share?
Many thanks
Brittany
Dear Brittany, it will depend on the antigen, which should stimulate CD8 T cells in your experimental settings. A huge quantity of variants of TCR may exist on CD8 T cells. So, unlikely anyone have CD8 cell line, which is necessary to you exactly. In addition, as a rule, immortalization of CD8 T cells results in the loss of CD8 coreceptor. I think, you have several options to detect CD8 cell priming: 1) Use TCR-transgenic mice with well known specificity of transgenic TCR. In this case you can detect CD8 T cell response by proliferation in MLR with stimulated DC and subsequent analysis by flow cytometry. 2) You can generate your own CD8 CTL cell lines specific to the antigen of interest. In this case you can detect responses by estimating CTL activity, IFN gamma production, etc. Unfortunately, CD8 CTL lines are unstable in vitro. So you will have only restricted time to perform experiments. 3) You can generate your own T cell hybridomas, using BWZ.36.CD8 alpha thymoma cells (developed by prof. Nilabh Shastri) as tumor partner for fusion with T cells, activated by the antigen of interest. Here, you can detect response by estimating NF-AT in LacZ assay or IL-2 production in CTLL assay. The stability of T cell hybridomas also leaves much to be desired, but you will have several months to do experiments. Good luck!
- Badges
- Science method