Hi,
I am using Balb/c spleen for DC and B6 spleen for T cell. Both them are isolated by milteny biotec kit (pan DC and pan T cells). Then I directly put them together for MLR. On day 5, I can see the T cell proliferation by CFSE staining. However, it only from CD8 T cell. Is it correct? Because I check the T cell, It has CD4 and CD8 two subsets on beginning and also on day 5, the naive T cell still contains CD4 and CD8. But just CD8 t cell get proliferation.
Thank you
Dear Dr Hsi-Ju Wei, proliferation of CD8+ cells is faster in comparison with CD4+ T cells. Then, you see predominance of CD8+ cells. If stimulate B6 T cells with DC from bm12 (A-beta(b) mutant mouse strain), you will see selective proliferation of CD4+ T cells. Against, if stimulate B6 T cells with DC from bm1 (K(b) mutant mouse strain), you will see proliferation of CD8+ T cells. Enclosed picture shows results of FACS analysis of 3 day MLR.
Hi Dmitry,
I understand if I am using the mutant mice on MHC complex for T cell in MLR. I just wonder can I see the CD4 and CD8 both proliferate in normal B6. Because naive T cell have CD4 and CD8 positive. But can they are activated and become blast, I only see CD8 positive. Should I do some modification to see the CD4? Here is my result. FL4=CD4, Percp=CD8, FITC=CFSE.
CD8 from:5% to 64%
CD4 from 3% to 9%
You should see proliferation of both CD4 and CD8 T cells. Enclosed picture shows this for naïve T cells and T cells from mice preliminary immunized with EL4 (H-2b) tumor cells.
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