Recently a technique for quantifying cytokine producing cells using the LSC was developed. Using the protocol originally developed for FISH spot counting, we were able to observe and quantify the characteristic staining morphology, and thereby sort out the true positive cells from autofluorescent and non-producer ligand binding target cells.
Cytokines are protein hormones that function as growth factors and regulators of cellular responses.
This family of secreted, regulatory glycoproteins contains more than 30 members. They are produced by lymphocytes, granulocytes, and macrophages but also by cells outside the immune system. They serve as important mediators in inflammatory and immune responses but are also implicated in the pathophysiology of various diseases. Most cytokines have a signal-sequence, regulated secretion pathway, and their proforms are often accumulated in the Golgi organelle. Human peripheral blood mononuclear cells were harvested and stained for IL-6 after 6 hours of LPS stimulation.