I'm replacing primary neutrophils with the neutrophil-like cell line HL-60 for immunological studies.
Do HL-60 cells, differentiated towards the granulocyte phenotype with retinoic acid or DMSO, secrete inflammatory mediators such as interleukins in response to LPS?
I'm unfamiliar with their expression of TLR4/MD2/CD14.
Hello, you may consider consulting this and other papers:
https://pubmed.ncbi.nlm.nih.gov/18291094/
Since TLR4, and capase-1 pathways are present, HL-60 should respond to LPS, though the strength of which may vary according to differentiation status.
THP-1 for example (as a model of differentiated macrophages), can be "optimally" differentiated with very low dose of PMA (5 ng/mL; Article Optimized THP-1 differentiation is required for the detectio...
) before LPS challenge and other manipulations. The doses and duration of PMA affect differentiation outcomes. I can imagine some similar scenarios for HL-60. A good way to go about it, may be first find some "optimal" conditions for differentiating the cells, based on methodolgy papers.If in fact, your differentiation method is new and improved (with strong reproducibility), you can actually compare the expression profiles changes before and after (IL-6, IL-1beta, NF-kB activation, iNOS, MPO, etc.). That itself could be a worthwhile way to establish a model.
https://www.hindawi.com/journals/mi/2017/6909415/
https://pubmed.ncbi.nlm.nih.gov/24746556/
https://pubmed.ncbi.nlm.nih.gov/11298604/
Good luck!
Most of the myeloid-derived leukemic cells usually respond to TLRs stmulation by phophorilating Nf-kB resulting in secretion of TNF-a. Are you interested in a particular activation pathway or just any kind of activation will be fine ? Additionally, why you give up primary neutrophils ? they respond much better than any other cell line
Dario Armando Leone Thank you. I will be looking at inflammatory pathway activation using intracellular phospho arrays and similarly profiling cytokine secretion. It would be good if they secrete IL-8 as I have an ELISA to determine an LPS dose response, for example. I'm no longer able to use primary neutrophils from blood due to the COVID-19 situation.
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