I want to reconstitute the mice with genetically modified CD4 cells after withdrawal of endogenous CD4 cells. Thus, since antibodies are quite stable, I guess antibody-mediated depletion is not the method of choice. Any suggestions?
would be nice have recipients with CD4 T-cells expressing DTR! don't know if this mouse exist.
or Ab mediated depletion based on allele differences like CD45.1/CD45.2 or Thy1.1/Thy1.2...
Perhaps you might lethally irradiate a set of mice and apart collect CD4+ lymphocytes from a second set of mice, using this time negative CD4+ selection or a combination of cell purification methods as ficoll or glass wool.
It might be too much work/money what I propose, but ideally you could try to use a Tamoxifen-induced-Cre system in combination with a CD4-CD8/CD3 -Cre construct (or whatever cell type you are interested and there is a tamoxifen inducible-promotor mice). It would take some backcrossing and optimizing the tamoxifen doses (injection or in water), but that would be a pretty good approach. hence you could tamoxifen-deplete the endogenous T cells and then perform either an adoptive transfer of your KO CD4s or even consider a BM-transfer +/- irradiation (irradiation is not absolutely necessary for reconstituting some compartments depending on the time frame). It all depends on what you want to actually test. Good luck! The DTR-CD4 approach mentioned by Maximiliano here is also a good idea.
In this case I think the sub-lethal irradiation followed by CD4 T cell transfer may be a viable option as long as your modified T cells are from the same strain. Of course this would be a chimeric T cell population and would depend on your modified T cells having no survival defects in a competitive setting. Since you are using BALB/c I would suggest using congenic Thy1.1 T cells so you can easily detect donor vs host T cells.
supporting Maximiliano´s comment: Depletion of CD90.1 expressing T cells in vivo with a mAb clone 19E12 is very efficient and specific. Thus, to reconstitute Thy1.1 congenic Balb/c recipient mice with your CD90.2 T cells of interest is in my opinion the most easy and fast approach (without the need of generating/importing new lines; the recipients and the mAb are commercially available)
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