Hi all. I am looking for an alternative for PBMC T-cells for performing the mixed lymphocyte reaction (MLR). I have found 1-2 references where Raji cells are used as the stimulator cells and Jurkat cells as the responder (Neuber et al 2006).
However, we do not have these two cell lines in our lab. I was wondering if anyone knew if alternative T-cell lines (such as HH cells) have been used in this technique?
Also does anyone know if any adherent lines (such as non-transfected fibroblasts) have been used as the APC (stimulator) in this type of reaction?
Thanks in advance,
Gary
Dear Gary, I have severe doubts that substitution of PBMC by established immortalized T cell lines in the MLR will be adequate. These cell lines have disrupted activation machinery and proliferate indefinitely. As a rule, adherent cell line are not professional APC. Then, you will have chance to see proliferation in your MLR only if your responder population contains clones of memory cells specific to HLA molecules on stimulators.
Hi Dmitry - thanks for taking the time to answer. You are correct in that these lines do proliferate indefinitely but appear (according to Neuber et al above and several other papers I have seen) to be applicable in measuring the immune-suppressive effects of mesenchymal stromal cells (MSC) in culture (where a decrease in proliferation compared to controls is observed in these cells). Another leukemic cell line (Karpas 299) has also been used in a similar way (alongside traditional MLR, Hill et al 2012) although in this case the Karpas cells were used as the "responder" (Treg) T-cells against normal T-effector cells (Teff) isolated from PBMCs (and they measured CD154 induction rather than proliferation).
I agree that cell lines cannot substitute for all of the different responses etc that you might get in a traditional MLR but it seems pretty clear that for certain specific aspects of the MLR (effects of factors on immune suppression by upregulation of Tregg-like cells and a corresponding reduction in proliferation) some lines can be used. However (as you implied) the functions and ploidy of cell lines compared to primary cells is pretty scrambled and thus perhaps only certain cell lines (such as Jurkat/Raj cells) can be used in this limited way.
My question was really to see if anyone had used cell lines (other than these two) to mimic some of the MLR aspects. We have HH cells (CRL-2015) and Jurkat E6 (but not the parental Jurkat line) in our lab and these were the two lines I was really wondering about (but failed to specify - mea culpa!).
Thanks again for taking the time to answer!
Gary