I am trying to stimulate oral epithelial cells for cytokine release. I have tried different endotoxins, but not able to get any results.
Any suggestions?
Hello Minati,
Do you use primary oral epithelial cells (OECs) from human donors, or do you use oral carcinoma-derived cell lines such as H413 or TR146?
The thing is, primary oral epithelial cells from human donors are naturally exposed to a variaty of different bacterial antigens, which are present in the oral mucosa and our food. These cells tolerate bacterial antigens, which are also present on commensal bacteria in our oral cavities, to avoid an over-reaction of the immune system. It will be difficult to get these cells to release cytokines.
I would suggest to use cell lines such as H413 or TR146 and different bacterial stimuli from both, gram+ and gram- bacteria.
In the following study, the authors used MV130, which is a cocktail of different heat-inactivated whole cell bacteria, to stimmulate H413, TR146 and also primary oral epithelial cells:
(Molero-Abraham et al. 2019) Human Oral Epithelial Cells Impair Bacteria-Mediated Maturation of Dendritic Cells and Render T Cells Unresponsive to Stimulation. Front Immunol. 2019 Jun 28;10:1434.
doi: 10.3389/fimmu.2019.01434
H413 and TR146 cells produced at least IL-6 and IL-8 after stimulation with MV130, whereas primary oral epithelial cells did not respond to MV130 stimulation.
I hope, this will help you.
Kind regards,
Alexander Gluschko
Dear Alexander,
Thank you so much .
I am using oral carcinoma derived cell line H400. I have tried different TLR agonists, most of them have not worked. Lately I had single success with Poly I.C. I still need to replicate and confirm the result. Thanks for the suggestions regarding bacterial stimuli. This is helpful. I am thinking of using heat killed E.coli.
Hopefully that works.
kind regards
Minati
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