Hi everybody
I am developing an adjuvant but it is in the very initial steps. I want to first test it in vitro on human cells. I do not know which cells should I choose. I know that dendritic cells (DCs) are the most important cells in antigen presenting but if I choose them the role of other cells (neutrophil, monocytes, NK, etc.) will be ignored. Human PBMC seems to be a good choice but the result of it may be very different to real environment of human subcutaneous or intramuscular where the final adjuvant is expected to use. Furthermore the majority of previous articles focused on DCs. So I am confusing. Could you please help me
You are right most of these experiments are performed using DC or langenhans cells. The skin DC (Langerhans cells) may be the best for your applications. Highly active in antigen presentation, but also difficult to isolate.
Without any further context it is difficult to confidently suggest a preference however I would be inclined toward a DC-based assay. One could do this by isolating the monocytes from the PBMCs and then promoting moDC differentiation using GM-CSF and IL-4. There are plenty of methods describing this in the literature and I think it would be more definitive/less ambiguous than using PBMCs. Just to be clear I am not an expert on handling human DCs - I primarily work with murine BMDCs.
Your experimental design will depend on the nature of your adjuvant. You would definitely want to use DCs to begin with. However, if your adjuvant is also expected to have direct influence on T/NK cells and neutrophils through TLR ligation or other mechanism, you should then use PBMC or DC-lymphocyte co-cullture system.
Thank all
I think I'd better to start with DC. Then I may try both of them
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