I want to study cytokine production upon S. aureus infection in vitro. I have the following concerns:
1. Which human/mouse cells are best suited ?
2. Which assay is best suited in terms of both feasibility and sensitivity ?
Looking forward to suggestions, please
I agree with Gina.
1. Probably a myelomonocytic/macrophage cell line is suitable for cytokine detection. Human cell lines you could use include THP-1, or U937, but you can also use monocyte-derived macrophages, differentiating monocytes with GM-CSF (M1) or M-CSF (M2). In mouse model, you can use J774 macrophage cell line, or primary peritoneal macrophages / bone-marrow derived macrophages.
2. The best way to detect any cytokine released is ELISA. Nevertheless, maybe you could also try to detect cytokine accumulation using intracellular staining, prior brefeldin A treatment, and acquisition by flow cytometry. It depends on the sensitivity of your system, and I usually prefere ELISA, because it gives you concentration of the molecule.
Hope it helps
Dear Sudip,
According to our studies, HIV-1 infection in vitro of monocyte-derived macrophages (MDM) might serve the very appropriate model you are looking for. Also, a number of cell lines, mostly Mono Mac, THP-1, U937, HL-60 have complemented the MDM system of infection providing useful information on the features of HIV-1 replication in MDM, as well as cytokines production. Thus macrophage activation was shown by TNF production and NF-κB activation upon HIV-1 infection.
Best wishes,
Ilya
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