Abstract
One of the most powerful tools in immunotoxicology is the assessment of cytokines, the proteins/peptides responsible for regulating a variety of processes including immunity, inflammation, apoptosis, and hematopoiesis. Cytokine production measurements offer outstanding information and may eventually substitute for other more laborious procedures in the assessment of immunotoxicity. The particular profile of cytokine production provides indeed important information regarding the nature of many immunotoxic responses.
Recent expansion in the knowledge of cytokine biology and the realization that cytokines play a role in human diseases have created a need for the precise assessment and accurate interpretation of their presence and activity in body fluids, tissues, and cells. Proper evaluation of cytokines requires attention to several technical details. Multi-cytokine analysis still needs to be standardized in terms of optimum source for analysis, protocols, and quality control issues, such as the use of reference standards and the expression of results.
Important practical details and considerations will be discussed in this chapter, including the source of the sample to be tested (circulating fluids or ex vivo/in vitro isolated cells); the potential effects of collection, processing, and storage of the results of the assays, as well as potential variables associated with the source material (matrix effects, relevance, inhibitory substances); and factors influencing the choice of assay used (bioassay, immunoassay, molecular biology technique, flow cytometry).
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References
House RV, Descotes J (2007) Cytokines in human health: immunotoxicology, pathology and clinical applications. Humana Press, Totawa, NJ
House RV (2001) Cytokine measurement techniques for assessing hypersensitivity. Toxicology 158:51–58
Whiteside TL (2002) Cytokine assays. BioTechniques 10:12–15
Vandebriel RJ, Van Loveren H, Meredith C (1998) Altered cytokine (receptor) mRNA expression as a tool in immunotoxicology. Toxicology 130:43–67
Overbergh L, Giulietti A, Valckx D et al (2003) The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression. J Biomol Tech 14:33–43
Jason J, Larned J (1997) Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods 207:3–22
Pala P, Hussell T, Openshaw PJ (2000) Flow cytometric measurement of intracellular cytokines. J Immunol Methods 243:107–124
Letsch A, Scheibenbogen C (2003) Quantification and characterization of specific T-cells by antigen-specific cytokine production using ELISPOT assay or intracellular cytokine staining. Methods 31:143–149
Robroeks CM, Jöbsis Q, Damoiseaux JG et al (2006) Cytokines in exhaled breath condensate of children with asthma and cystic fibrosis. Ann Allergy Asthma Immunol 96:349–355
Winkler O, Hadnagy W, Idel H (2001) Cytokines detectable in saliva of children as appropriate markers of local immunity of the oral cavity-an approach for the use in air pollution studies. Int J Hyg Environ Health 204:181–184
Simpson JL, Wood LG, Gibson PG (2005) Inflammatory mediators in exhaled breath, induced sputum and saliva. Clin Exp Allergy 35:1180–1185
Whicher J, Ingham E (1990) Cytokine measurements in body fluids. Eur Cytokine Netw 1:239–243
Ryan CA, Gerberick GF, Gildea LA et al (2005) Interactions of contact allergens with dendritic cells: opportunities and challenges for the development of novel approaches to hazard assessment. Toxicol Sci 88:4–11
Mitschik S, Schierl R, Nowak D et al (2008) Effects of particulate matter on cytokine production in vitro: a comparative analysis of published studies. Inhal Toxicol 20:399–414
Jeurink PV, Vissers YM, Rappard B et al (2008) T cell responses in fresh and cryopreserved peripheral blood mononuclear cells: kinetics of cell viability, cellular subsets, proliferation, and cytokine production. Cryobiology 57:91–103
Kreher CR, Dittrich MT, Guerkov R et al (2003) CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays. J Immunol Methods 278:79–93
Maecker HT, Moon J, Bhatia S et al (2005) Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT. BMC Immunol 18:6–17
Arora SK (2002) Analysis of intracellular cytokines using flowcytometry. Meth Cell Sci 24:37–40
Ghanekar SA, Maecker HT (2003) Cytokine flow cytometry: multiparametric approach to immune function analysis. Cytotherapy 5:1–6
Maecker HT, Rinfret A, D'Souza P et al (2005) Standardization of cytokine flow cytometry assays. BMC Immunol 6:13
Meager A (2006) Measurement of cytokines by bioassays: theory and application. Methods 38:237–252
Mire-Sluis AR, Page L, Thorpe R (1995) Quantitative cell line based bioassays for human cytokines. J Immunol Methods 187:191–199
House RV (1999) Cytokine bioassays: an overview. Dev Biol Stand 97:13–19
Blumberg DD (1987) Creating a ribonuclease-free environment. Methods Enzymol 152:20–24
Suni MA, Maina VC, Maecker HT (2005) Ex vivo analysis of T-cell function. Curr Opin Immunol 17:434–440
House RV (2002) Preclinical immunotoxicity assessment of cytokine therapeutics. In: Thomas JA, Fuchs RL (eds) Biotechnology and safety assessment, 3rd edn. Academic Press, San Diego
Heney D, Whicher JT (1995) Factors affecting the measurement of cytokines in biological fluids: implications for their clinical measurement. Ann Clin Biochem 32:358–368
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Corsini, E., House, R.V. (2018). Evaluating Cytokines in Immunotoxicity Testing. In: DeWitt, J., Rockwell, C., Bowman, C. (eds) Immunotoxicity Testing. Methods in Molecular Biology, vol 1803. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8549-4_18
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DOI: https://doi.org/10.1007/978-1-4939-8549-4_18
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