Hi all,
We have germline KO, CD4cre KO and Foxp3cre KO mice of the same gene. But when we sorted out naive CD4 T cells using CD4+, CD62L+ and CD25-, and subjected these cells to iTreg differentiation. We could find that germline KO naive CD4 T cell have higher iTreg than WT. But we could not find the same phenotype on CD4cre and Foxp3cre KO. We checked mRNA of our gene of interest, the cre system works fine. Does any one know why?
Hi,
since you observed a higher iTreg polarization in germline KO naive CD4+ T cells compared to WT cells, and T cell or Treg specific knockout of the gene could not phenocopy what you see in germline KO, it is indicative of other cell types to be responsible for the iTreg differentiation. ie. The deficiency of the gene of your interest on other cell types induced higher iTreg differentiation.
Best
Hi Yi,
Maybe I didn't state it clear. I sorted out naive CD4+ T cells and put them in iTreg polarization condition (TGFb and IL-2 with plate-bound anti-CD3/CD28). There is no other cells in the system. In terms of our GOI(gene of interest), both germline and CD4cre KO cells are deficient of our GOI at onset of experiment. So I can't understand why they show different results. Is it because something happened before CD4 expressed during naive CD4 T cell development? That is why I'm asking whether naive CD4 T cells are identical if they have the same genomic background. For example, if we KO some gene in mTEC or cTEC or DC etc.. Do the naive CD4 T cells isolated from these mice are identical?
Hi Jian,
I thought it was co culture system. Sorry for the misunderstanding and thank you for the details on your concern. I searched online resources on the factors that might affect naive CD4+T cells. I didn't find any clues about the DC/TEC on the instruction of naive T cell property. However, I do find the research demonstrating that TCR Repertoires of CD4 Naïve T Cells Change Properties With Aging (Article The Changing Landscape of Naive T Cell Receptor Repertoire W...
;Article Transcription factor networks in aged naïve CD4 T cells bias...
). I'm not sure whether you GOI is a determinant for aging or not. If it's positive, it might explain the observation in our case. If you find other evidences linking DC and naive T cell training, I would really appreciate it.Best
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