I am using PBMC from normal donors, I have non-stimulated controls and 24hr stimulated samples with 1:1 Dynabeads. In the non stimulated samples, my CD4 CD8 gating looks normal (only 2 pop: either CD4+ or CD8+). However, in the stimulated samples I have a pop of CD8+ and a clear and large double positive population CD4+ and CD8+ and no CD4+ only population.
Has anyone ever seen this before? Is it something that happen with this kind of stimulation?
Thank you
Hi Francesca
Did you use any antigen or cytokine to stimulate the cells? CD4+ CD8+ cells can be generated in response to certain antigens or cytokine composition. It would be interesting to measure the cytokine profile (TGF beta, IFN gamma...). You may find this article useful
Best wishes
Siva
Article CD4+/CD8+ double-positive T cells: More than just a developm...
Thank you. It turns out, after several trials and re-running the experiment and by looking at the FMOs that I really did not have any significant CD8 staining. I am using anti human CD8 AF700 from Invitrogen, clone RPA-T8. I compared it to several other anti CD8 antibodies in other colors and sure enough, it was not providing any staining. So what I thought were double pos CD4 and 8, were really single CD4+ cells....
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