I am testing a bacterial mutant in pneumonia model and also wanted to check for inflammation. Apart from microscopic examination using HE staining, I wanted to quantify inflammatory cytokines in the lung (also in lung cell lines in vitro). What are the major cytokines, which can quantified as hallmarks of inflammation in lung ?
Hey Bro!! i think you can quantify IL4, IFN-gamma, IL-10, IL-6. But before that you can try to figure out what is the response type.. is it Th1 or Th2. basing on that you can select the cytokines you want to quantify. i had recently come across a paper published in Nature-2013 which states involvement of Th-17 family of cytokines including IL17. i think you can refer that...(http://www.nature.com/emi/journal/v2/n9/full/emi201358a.html)
Hey Sudip, our group normally quantifies IL-1beta, IL-6, TNF-alpha and IL-12, but, as our colleague said, you should to figure what type of response your bacteria will induce Th1, Th2 or even Th17. Good lucky to you!
Hi Sudip,
As others have mentioned, you will have to figure out the kind of inflammatory response you expect to see in your model (Th1/Th2/Th17). In case you are starting from scratch I would suggest running a multiplex assay (cytokine/chemokine) with representative targets from each pathway along with pro-inflammatory cytokines like TNF-a and IL-8. This might help you narrow down a specific pathway that you can further explore.
Also, what type of cells are you looking at in-vitro (epithelial, fibroblast, smooth muscle etc)? we use esophageal epithelial cells and they produce tonnes of IL-8 on stimulation with the TLR-3 synthetic ligand poly (I:C) (viral dsRNA mimetic)
Hi Sudip
I would suggest the following important cytokines TNF-α, IL-1β, IL-10 and HMG-1. Also you can try TLR-3 and IL-8 as well.
Good luck.
Sudip, IL-8, RANTES, MCP-1, IFN-g, and TNF-a might be worth looking at in these cells. If you are seeing eosinophilic infiltration in you tissue sections, you might also want to look at some Th2 cytokines. Also, starting with running qPCRs for these targets might help you save money and time since these cytokine multiplex assays are usually expensive. I would pick my targets based on what I am seeing at the gene expression level.
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