I am trying to simulate an in vivo situation in host macrophages by adding a recombinant protein that is expressed in high amounts.Can i do cytokine upregulation studies without adding PMA ?Will it give any useful information on the real situation in the host?.Please see this paper below which talks of using LPS alone or Shigella toxin to look for cytokine upregulation-directly on THP-1 cells
How is your recombinant protein produced? From what organism is it derived? In my opinion it will not model the host response because you need to consider that THP-1 cells are very different from primary cells and the host response will definitely be the response of multiple cell types. Also, the purified recombinant protein will likely be structurally similar but have different or no post translational modifications. In the situation you describe you would only be modeling how THP-1 cells respond to a recombinant protein, potentially very interesting data, though this does not model the in vivo situation.
THP1 cells can be a useful starting point for your studies. THP1 cells are monocytic and largely non-adherent. PMA treatment is necessary for the cells to become macrophage-like. Some features of THP1 responses accurately reflect ex vivo macrophage responses. BM or PB derived macrophages should be compared. In vivo responses (eg serum cytokines responses) involve multiple cell types and thus don’t reflect simply the response of macrophages.
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