Hello,

I'm staining thawed human PBMC with a Th panel (live/dead, CD3, CD4, CD25, CD127, CXCR3, CCR4, CCR6, CCR10). I usually gate live cells > CD3+ CD4+ T cells > CD25+ CD127- Tregs, then I define the "NOT Tregs" population in which I look at the chemokine receptors expression. I expected to see CXCR3+ CCR4- cells (i.e. Th1 and/or Th17.1, according to the expression of CCR6), as I can see this population very clean in fresh PBMCs. But in thawed PBMCs it turns that CXCR3+ cells are all CCR4+.

Do you think that cyropreservation/thawing could drive this CCR4 expression on PBMCs? What should I do to overcome this problem?

Thanks in advance for your advice!