I would like to know if my endosomal TLRs (e.g. TLR7/8/9/3) are activated. Are there endosomal TLRs specific cytokine production in Macrophage? Can someone tell me which cytokines are produced after activation of above TLRs (e.g. TLR7) but not any with other TLRs (e.g. TLR4). Or is there other way to know the activation of endosomal TLRs in macrophages.
Any help is appreciated.
Thank you
Most TLRs that you noted activate pro-inflammatory response that can be assessed by activation of NF-kB target genes and IRFs target genes.
Also, you can perform a western blot to check the entire signalling cascade of TLRs, including MAPK, IRF and NF-kB target genes.
You could try to include ibhibitors of endosomal maturation like chloroquine or bafilomycin A or inhibitors of endocytosis like monodansylcadaverie in your assays.
see also:
- The Toll-like receptor 7 (TLR7)-specific stimulus loxoribine uncovers a strong relationship within the TLR7, 8 and 9 subfamily
- CpG-DNA-specific activation of antigen-presenting cells requires stress kinase activity and is preceded by non-specific endocytosis and endosomal maturation
- Bacterial CpG-DNA and lipopolysaccharides activate Toll-like receptors at distinct cellular compartments.
Thank you Dr. Divaker Choubey and Dr. Lavinia Romera. I got some direction from your answers.
Thank you Dr. Parviz Ahmad-Nejad for suggesting reading materials. I like the idea of including inhibitor of endosomal maturation.
Again, thank you all for your time and writing answers.