Cytokine Production by Stimulated Immune Cells
Isolated whole blood can be stimulated in culture medium to produce various cytokines by incubation for 24 h at 37oC with either 5 μg/ml E. coli lipopolysaccharide (LPS) or 10 μg/ml phytohemagglutinin (PHA) as indicated below. Cytokine concentrations released into the medium can be measured either directly by standard ELISA techniques or indirectly by assaying for a specific biological function of the particular cytokine in question, as illustrated in the preceding section for example.
Monocytes/macrophages in
whole blood cultures (refs. 1, 2, 6).
- LPS primarily stimulates IL-1β, IL-1ra, IL-6, IL-10, IL-12, and TNF-α production.
- LPS plus PHA stimulates greater production of IL-6 and TNF-α and reduced production of IL-1β as compared to LPS alone (1).
T cells in whole blood cultures (refs. 1, 3, 4).
- PHA primarily stimulates IL-2, IL-4, IFN-γ, IL-17, and GM-CSF.
- PHA plus LPS stimulates greater production of IFN-γ but only slightly greater production of IL-2 and GM-CSF, as compared to PHA alone.
- LPS alone does not stimulate
production of IL-2, IFN-γ or GM-CSF from whole blood cultures (1).
NK cells in PBL cultures (ref. 5).
- Anti-CD2 (as opposed to anti-CD3; see ref. 5) primarily stimulates TGF-β production.
References:
- D. De Groote, P. F. Zangerle, Y. Gevaert, M. F. Fassotte, Y. Beguin, F. Noizat-Pirenne, J. Pirenne, R. Gathy, M. Lopez, I. Dehart, D. Igot, M. Baudrihaye, D. Delacroix, and P. Franchimont. Direct stimulation of cytokines (IL-lβ, TNF-α, IL-6, IL-2, IFN-γ and GM-CSF) in whole blood. I. Comparison with isolated PBMC stimulation. Cytokine 4:239-48 (1992).
- E. F. Schippers, C. van 't Veer, S. van Voorden, C. A. E. Martina, T. W. J. Huizinga, S. le Cessie, and J. T. van Dissel. IL-10 and toll-like receptor-4 polymorphisms and the in vivo and ex vivo response to endotoxin. . Cytokine 29:215-28 (2005).
- X. Paliard, R. de waal Malefijt, H. Yssel, D. Blanchard, I. Chretien, J. Abrams, J. de Vries, and H. Spits. Simultaneous Inroduction of IL-2, IL-4, and IFN-γ by activated human CD4+ and CD8+ T cell clones. J. Immunology 141:849-55 (1988).
- K. Kurasawa, K. Hirose, H. Sano,
H. Endo, H. Shinkai, Y. Nawata, K. Takabayashi, and I. Iwamoto. Increased interleukin-17 production in patients with systemic sclerosis. Arthritis Rheumatism 43:2455-63 (200O).
- J. D Gray, M. Hirokawa, K. Ohtsuka, and D. A. Horwitz. Generation of an inhibitory circuit involving CD8+ T cells, IL-2, and NK cell-derived TGF-β: Contrasting effects of anti-CD2 and anti-CD3. J. Immunology 160:2248-54 (1998).
- J. L. Nerad, J. K. Griffiths, J. W.M. Van der Meer, S. Endres, D. D. Poutsiaka, G. T. Keusch, M. Bennish, M. A. Salam,
C. A. Dinarello, and J. G. Cannon. Interleukin-lβ (IL-lβ), IL-1 receptor antagonist, and TNF-α production in whole blood TGF-β. J. Leukocyte Biol. 57:687-695, 1992).