Sooner or later they will kill them. Since monocytes give rise to a variety of macrophages (with more reperatory or stimulatory capacities) it strongly is influenced by your culture conditions and stimulations. In vivo CD8+T-cells need CD4+ T cell help. Especially when they r antigen inexperienced. I don't know what kind of cd8+t-cells you'll use.
Meybe you'll need a restimulation (after 1 week e.g.) and the addition of cytokines like IL-2 and/or IL-15 to see a strong and potent killing. I'm no real expert, though
I got some experiences with dendritic cells derived from monocytes and CD8+T-cells in vitro.
Hope this helps
Regards
Robert
The proportion of T-cells reactive against that specific HLA is going to be very small. As Robert said, you would have to undergo a complete in-vitro antigen-dependent expansion process, in order to grow the alloreactive compartment up to a meaningful proportion of the total T-cell pool. It's very difficult to do this on purpose, and certainly takes a long time. If you see your monocytes dying in the short run, it's probably due to a different mechanism.
Thank you. for your input. I would add that these T cells are specific for a certain HLA-restricted antigen and have been expanded in cultture for at least 2 weeks to enrich the population for CD8+ T cells. Would such an expansion protocol sufficiently generate T cells that are capable of an allogenic reaction against HLA-mismatched monocytes? If not, could you comment on what alternative mechanism is responsible for this observed "allogenic rejection" of monocytes? Thanks in advance.
Now I'm a little confused. Do you have already CD8+ T Cells primed for this specific HLA subtype? (primary and or restimulation!?) if so, they should kill your monocytes really fast, in my opinion.
The CD8+ T cells are primed for a specific MHC Class complex (consisting of a specific peptide). This complex is presented by a target cell line - that is not the monocyte. The monocyte is a third cell type that is artificially added to the 3-cell co-culture. The objective is to investigate the effect of monocytes on the interaction between the primed CD8+ T cells and the MHC:peptide-presenting target cell line (where TCR binding is peptide specific and restricted to that particular MHC Class). I observed that the effect seen with autologous monocytes was, instead, more short-lived when we used allogenic monocytes. Does this clarify? Please let me know where I can further clarify. Thanks Robert!
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