Hello all,
As I know, IL-8 can triggers neutrophil degranulation and therefore induces the release of MPO into the culture supernatant. However, I could not find any study investigating the direct effect of IL-8 on MPO production ex vivo.
Is there anyone has tested the effect of IL-8 on MPO release from neutrophils? How is the phenomenon?
Thank you very much for your information!
I'm confused by your use of the word "production". Neutrophils synthesise all their MPO and package it into azurophilic/primary granules while still in the bone marrow. There will be no synthesis ex vivo, only release (degranulation) of pre-synthesised enzyme.
Don't think that anyone ever has added IL-8 to granulocytes and measure MPO. It should increase, but no idea. Guess one of the problems is that IL-8 can signal through a variety of signalling cascades, which make it difficult to block one by siRNA for example, without affecting the others.
Hi Clark and Werling,
Yes I meant "release". I expected that IL-8 could induce the degranulation of neutrophils, therefore increase the concentration of MPO in the culture supernatant. However, concentration of MPO was decreased when I treated neutrophils with recombinant IL-8 (I tested more than 10 independent times so I am quite sure about the data).
It's not consistent with what I have read so I would like to ask for your experience if you also tested the effect of IL-8.
If you know any report about this issue, please let me know.
Thank you so much.
Dear Pam
MPO activity of neutrophils can change with the stimulii. Although, this enzyme is in the azurophylic granules its activity has been seen to change according the inflammatory microenvironment for different in vitro studies (Piwowarski et al, 2014; Dalboni et al, 2013). A very good review about it is: http://www.ncbi.nlm.nih.gov/pubmed/23066164
You should analyze cell viability in the neutrophils stimulated with IL8. If you measures MPO through its enzyme activity, this can be reduced whether cells are not alive and whether the time of your culture is long. If you determines MPO concentration according with the expression of protein (western blot) you should not find decrease in response to IL8
Thank you for your suggestion Gasparoto.
Hi Dios, as you said i think IL-8 treatment could affect neutrophil survival. Do you have any idea about this issue? Thank you again
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