I would like to adoptively transfer OT-I T cells to immune-intact mice and examine tissue later by flow cytometry. Unfortunately, our OT-I T cells are CD45.2 and I do not think we can afford several cages of CD45.1 mice so that I can identify the T cells by a congenic marker. Antibodies are cheaper and more reliable than tetramers, so I am considering using double staining for V alpha 2 and V beta 5. Is this acceptable?
Thanks for the input Jie! Unfortunately, I'm tracking a memory response, so I don't think dyeing the donor cells will last long enough.
If it is not an urgent experiment, you can get one male CD45.1.1 mouse and set up a breeding with CD45.2.2 females. The F1 mice with CD45.1.2 are equally good.
I agree with Jie that there will be some endogenous Va2Vb5 non-OT-I cells. But it really depends on the experiment and read-out whether this bothers you or not. By staining Va2Vb5 you will get more or less all transferred OTI (memory) cells, thus for functional tests afterwards this should be ok and the "dilution" with endogenous Va2Vb5 would not harm too much. When you want to count the cells in different locations this will be less precise.
But may be staining Va2Vb5 is not a bad idea at all beside tracking markers. Recently there was a publication in NI dealing with the loss of the OT-I transgene:
Spontaneous partial loss of the OT-I transgene - p471
Gretchen Harms Pritchard, Eric W Cross, Marjorie Strobel, Stephen C Jameson, Ross M Kedl, Kristin A Hogquist & Christopher A Hunter; doi:10.1038/ni.3411