Abstract
Substantial interest has been generated by the potential roles of the cytokines in health and disease (1). This has prompted considerable investigation into how these mediators are regulated to answer such basic questions as which stimuli initiate transcription and what factors are responsible for inhibiting secretion. This has resulted in elegant studies that have begun to define the intracellular-signaling pathways responsible for the upregulation of cytokines (2,3). Many of these studies have been done with cultures of cell lines derived from cancers or primary cultures of isolated fibroblasts, endothelial cells, or isolated mononuclear cells. Whereas these studies have provided substantial insight, they may be limited in their scope because they do not include all cell-cell or cell-protein interactions that take place in vivo. Other studies have used endotoxin injection into normal human volunteers to study the upregulation of cytokines (4,5). These studies with the normal volunteers provide precise information about the kinetics of cytokine production, but they are difficult to perform and very expensive. The whole-blood model serves as a useful bridge between using normal volunteers and isolated peripheral blood mononuclear cells. For critically ill patients it would be impossible to perform endotoxin infusion studies, and it would even be difficult to conduct these types of studies in chronically ill patients. Whole blood may also be used to study the immune responses of such patients in an attempt to determine how their cytokine regulation differs from normal individuals.
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References
Remick, D. G. and Friedland, J. S., eds. (1997) Cytokines in Health and Disease, 2 ed. Marcel Dekker, New York, NY.
Remick, D. G. (1995) Applied molecular biology of sepsis. J. Crit. Care 10, 198ā212.
OāShea, J. J., Notarangelo, L. D., Johnston, J. A., and Candotti, F. (1997) Advances in the understanding of cytokine signal transduction: the role of Jaks and STATs in immunoregulation and the pathogenesis of immunodeficiency. J. Clin. Immunol. 17, 431ā447.
Suffredini, A. F., Reda, D., Banks, S. M., Tropea, M., Agosti, J. M., and Miller, R. (1995) Effects of recombinant dimeric TNF receptor on human inflammatory responses following intravenous endotoxin administration. J. Immunol. 155, 5038ā5045.
Martich, G. D., Danner, R. L., Ceska, M., and Suffredini, A. F. (1991) Detection of interleukin 8 and tumor necrosis factor in normal humans after intravenous endotoxin: the effect of antiinflammatory agents. J. Exp. Med. 173, 1021ā1024.
Kasahara, K., Strieter, R. M., Chensue, S. W., Standiford, T. J., and Kunkel, S. L. (1991) Mononuclear cell adherence induces neutrophil chemotactic factor/interleukin-8 gene expression. J. Leukocyte Biol. 50, 287ā295.
Doherty, J. F., Golden, M. H., Remick, D. G., and Griffin, G. E. (1994) Production of interleukin-6 and tumour necrosis factor-alpha in vitro is reduced in whole blood of severely malnourished children. Clin. Sci. (Colch). 86, 347ā351.
Kremer, J. P., Jarrar, D., Steckholzer, U., and Ertel, W. (1996) Interleukin-1, ā6 and tumor necrosis factor-alpha release is down-regulated in whole blood from septic patients. Acta Haematologica 95, 268ā273.
van der Poll, T., Coyle, S. M., Moldawer, L. L., and Lowry, S. F. (1996) Changes in endotoxin-induced cytokine production by whole blood after in vivo exposure of normal humans to endotoxin. J. Infec. Dis. 174, 1356ā1360.
Mitov, I. G., Kropec, A., Benzing, A., Just, H., Garotta, G., Galanos, C., and Freudenberg, M. (1997) Differential cytokine production in stimulated blood cultures from intensive care patients with bacterial infections. Infection 25, 206ā212.
Friedland, J. S., Hartley, J. C., Hartley, C. G., Shattock, R. J., and Griffin, G. E. (1996) Cytokine secretion in vivo and ex vivo following chemotherapy of Myco-bacterium tuberculosis infection. Trans. Royal So. Trop. Med. Hygiene 90, 199ā203.
Majetschak, M., Flach, R., Heukamp, T., Jennissen, V., Obertacke, U., Neudeck, F., Schmit-Neuerburg, K. P., and Schade, F. U. (1997) Regulation of whole blood tumor necrosis factor production upon endotoxin stimulation after severe blunt trauma. J. Trauma 43, 880ā887.
Dahlke, E., Schlag, R., Langenmayer, I., Frankenberger, M., Kafferlein, E., Subkowski, T., Emmerich, B., and Ziegler-Heitbrock, H. W. (1995) Decreased production of TNF and IL-6 in whole blood of CLL patients. Am. J. Hematol. 49, 76ā82.
Fischer, J. R., Schindel, M., Stein, N., Lahm, H., Gallati, H., Krammer, P. H., and Drings, P. (1995) Selective suppression of cytokine secretion in patients with small-cell lung cancer. Ann. Oncol. 6, 921ā926.
Elsasser-Beile, U., von Kleist, S., Stahle, W., Schurhammer-Fuhrmann, C., Monting, J. S., and Gallati, H. (1993) Cytokine levels in whole blood cell cultures as parameters of the cellular immunologic activity in patients with malignant melanoma and basal cell carcinoma. Cancer 71, 231ā236.
Elsasser-Beile, U., von Kleist, S., Fischer, R., and Monting, J. S. (1992) Impaired cytokine production in whole blood cell cultures from patients with colorectal carcinomas as compared to benign colorectal tumors and controls. J. Clin. Lab. Anal. 6, 311ā314.
Zangerle, P. F., De Groote, D., Lopez, M., Meuleman, R. J., Vrindts, Y., Fauchet, F., Dehart, I., Jadoul, M., Radoux, D., and Franchimont, P. (1992) Direct stimulation of cytokines (IL-1 beta, TNF-alpha, IL-6, IL-2, IFN-gamma and GM-CSF) in whole blood: II. Application to rheumatoid arthritis and osteoarthritis. Cytokine 4, 568ā575.
Chofflon, M., Juillard, C., Gauthier, G., and Grau, G. E. (1991) Correlation between in vitro cytokine production and clinical evolution of multiple sclerosis patients. Schweiz. Arch. Neurol. Psychiatr. 142, 107ā112.
Chofflon, M., Roth, S., Juillard, C., Paunier, A. M., Juillard, P., Degroote, D., and Grau, G. E. (1997) Tumor necrosis factor production capacity as a potentially useful parameter to monitor disease activity in multiple sclerosis. Eur. Cytokine Network 8, 253ā257.
Hautecoeur, P., Forzy, G., Gallois, P., Demirbilek, V., and Feugas, O. (1997) Variations of IL2, IL6, TNF alpha plasmatic levels in relapsing remitting multiple sclerosis. Acta Neurologica Belgica 97, 240ā243.
Evans, C. A., Jellis, J., Hughes, S. P., Remick, D. G., and Friedland, J. S. (1998) Tumor necrosis factor-alpha, interleukin-6, and interleukin-8 secretion and the acute-phase response in patients with bacterial and tuberculous osteomyelitis. J. Infect. Dis. 177, 1582ā1587.
van der Poll, T., Jansen, J., Endert, E., Sauerwein, H. P., and van Deventer, S. J. (1994) Noradrenaline inhibits lipopolysaccharide-induced tumor necrosis factor and interleukin 6 production in human whole blood. Infec. Immun. 62, 2046ā2050.
van der poll, T. and Lowry, S. F. (1997) Lipopolysaccharide-induced interleukin 8 production by human whole blood is enhanced by epinephrine and inhibited by hydrocortisone. Infect. Immun. 65, 2378ā2381.
Hartman, D. A., Ochalski, S. J., and Carlson, R. P. (1995) The effects of anti-inflammatory and antiallergic drugs on cytokine release after stimulation of human whole blood by lipopolysaccharide and zymosan A. Inflamm. Res. 44, 269ā274.
Chan, B., Kalabalikis, P., Klein, N., Heyderman, R., and Levin, M. (1996) Assessment of the effect of candidate anti-inflammatory treatments on the interaction between meningococci and inflammatory cells in vitro in a whole blood model. Biotherapy 9, 221ā228.
Matsumoto, N., Aze, Y., Akimoto, A., and Tujita, T. (1998) ONO-4007, an anti-tumor lipid A analog, induces tumor necrosis factor-alpha production by human monocytes only under primed state: different effects of ONO-4007 and lipopolysaccharide on cytokine production. J. Pharmacol. Exp. Ther. 284, 189ā195.
Redl, H., Bahrami, S., Leichtfried, G., and Schlag, G. (1992) Solving one of the problems of in vitro cytokine production and endotoxin measurement in blood [letter]. Clin. Chim. Acta. 205, 149ā150.
DeForge, L. E., Tracey, D. E., Kenney, J. S., and Remick, D. G. (1992) Interleukin-1 receptor antagonist protein inhibits interleukin-8 expression in lipopolysaccharide-stimulated human whole blood. Am. J. Pathol. 140, 1045ā1054.
DeForge, L. E. and Remick, D. G. (1991) Kinetics of TNF, IL-6, and IL-8 gene expression in LPS-stimulated human whole blood. Biochem. Biophys. Res. Comm. 174, 18ā24.
DeForge, L. E., Kenney, J. S., Jones, M. L., Warren, J. S., and Remick, D. G. (1992) Biphasic production of IL-8 in lipopolysaccharide (LPS)-stimulated human whole blood. Separation of LPS-and cytokine-stimulated components using anti-tumor necrosis factor and anti-IL-1 antibodies. J. Immunol. 148, 2133ā2141.
DeForge, L. E., Fantone, J. C., Kenney, J. S., and Remick, D. G. (1992) Oxygen radical scavengers selectively inhibit interleukin 8 production in human whole blood. J. Clin. Invest. 90, 2123ā2129.
DeForge, L. E., Preston, A. M., Takeuchi, E., Kenney, J., Boxer, L. A., and Remick, D. G. (1993) Regulation of interleukin 8 gene expression by oxidant stress. J. Biol. Chem. 268, 12ā13.
Cale, D. R. and Remick, D. G. (1998) Low molecular weight heparin is associated with greater cytokine production in stimulated whole blood model. Shock 10, 192ā197.
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Remick, D.G., Newcomb, D.E., Friedland, J.S. (2000). Whole-Blood Assays for Cytokine Production. In: Evans, T.J. (eds) Septic Shock Methods and Protocols. Methods in Molecular Medicineā¢, vol 36. Humana Press. https://doi.org/10.1385/1-59259-216-3:101
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DOI: https://doi.org/10.1385/1-59259-216-3:101
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