If one does not purify cells from whole blood but stimulates whole blood, which cell types are stimulated by LPS ex vivo? Apart from just looking at the stimulatory effects of LPS, I am trying to understand what may be the relative contribution of each cell-subtype from acute lps-stimulation.
It depends on the time point. In the immediate, it is any cell that expresses TLR-4, which is mainly monocytes and granulocytes, with some on B cells. But almost as soon as the primary LPS stimulated cells activate, they in turn cause bystander acitvation of all the other cells.
So in whole blood LPS stimulation, there will be induction of all sort of cytokines at 24 hours.
Shen-An Hwang Thanks! Would you have any available literature on this based on your experience? That would be really useful. I am interested in kinetics
You might not find too much on whole blood, because most people don't do whole blood cultures. Most of the studies would be on purified macrophages/monocytes of either human or mouse origin, or human PBMCs. Mostly the former because TLR-4 is highly expressed on macrophages.
Most of my work with LPS were on macrophages/DCs, PBMCs, and the few experiments thta I've done where I took samples at 1 or 4 hours after LPS treatment (100ng/mL)... you see the obvious increases in cytokines (TNF-a, IL-6, IL-1b, IL-12p40, IL-10). And the only time I've ever did a whole blood culture with LPS
Check out these two:
1. DeForge and Remick. 1991. Kinetics of TNF, IL-6 and IL-8 gene expression in lps-stimulated human whole blood.
2. Huang et al. 2001.The plasticity of dendritic cell responses to pathogens and their components.
Emily Cornelius Ruhs Thanks! I do have those papers. Was wondering about contribution of cells types/ whole blood towards each of those cytokines.
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