Abstract
In order to get the full picture of the cytokine response generated by stimulation of various pathogens and superantigens in particular it is of crucial importance to collect data at various timepoints after stimulation. It is clear from those experiments performed that the production of the inflammatory and regulatory cytokines studied, differs in peak expression point and even cellular source depending on the time point studied (1,2). One other aspect of the importance of kinetics is that when one studies the cytokine profile of a human derived specimen, e.g., peripheral blood mononuclear cells (PBMC) the response varies between different donors both in number of cytokines produced, level of production as well as the kinetic of the response.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Björk, L., Andersson, J., Ceska M., and Andersson, U. (1992) Endotoxin and Staphylococcus aureus enterotoxin a induced different patterns of cytokines. Cytokine 4, 513–519.
Andersson, J., Nagy, S., Björk, L., Abrams, J., Holm, S., and Andersson, U. (1992) Bacterial toxin induced cytokine production studied at the single cell level. Immunol. Rev. 127, 69–96.
Andersson, U., Laskay, T., Andersson, J., and DeLey, M. (1986) Phenotypic characterization of single gamma-interferon producing cells after OKT3 antibody activation. Eur. J. Immunol. 16, 1457–1460.
Sander, B., Andersson, J., and Andersson, U. (1991) Assessment of cytokines by immuno.fluorescence and the paraformaldehyde/saponin procedure. Immunol. Rev. 119, 65–93.
Andersson, J., Abrams, J., Björk, L., Funa, K., Litton, M., Ågren, K., and Andersson, U. (1994) Concomitant in vivo production of 19 different cytokines in human tonsils. Immunology 83, 16–24.
Björk, L., Fehniger, T. E., Andersson, U., and Andersson, J. (1996) Computerised assessment of production of multiple human cytokines at the single cell level using image analysis. J. Leukocyte Biol. 59, 287–295.
Björk, L., Andersson, U., Chauvet, J.-M, Skansén-Saphir, U., and Andersson, J. (1994) Quantification of superantigen induced IFN-? production by computerised image analysis: inhibition of cytokine production and blast transformation by pooled human IgG. J. Immunol. Methods 175, 201–213.
Assenmacher, M., Schmitz, J., and Radbruch, A. (1994) Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 and interferon-? and interleukin-4 expressing cells. Eur. J. Immunol. 24, 1097–1101.
Jung, T., Schauer, U., Heusser, C., Neumann, C., and Rieger, C. (1993) Detection of intracellular cytokines by flow cytometry. J. Immunol. Methods 159, 197–207.
Ferrick, D. A., Schrenzel, M. D., Mulvania, T., Hsieh, B., Ferlin, W. G., and Lepper, H. (1995) Differential production of interferon-? and interleukin-4 in response to Th1-and Th2-stimulating pathogens by γ▽ T cells in vivo. Nature 373, 255–257.
Prussin, C. and Metcalf, D. D. (1995) Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J. Immunol. Methods 188, 117–128.
Henter, J.-I., Söder, O., and Andersson, U. (1988) Identification of individual tumor necrosis factor/cachectin-producing cells after lipopolysaccharide induction. Eur. J. Immunol. 18, 983–988.
Openshaw, P., Murphy, E. E., Hosken, N. A., Maino, V., Davis, K., Murphy, K., and O’Garra, A. (1995) Heterogeneity of intracellular cytokine synthesis at the single cell level in polarized T helper 1 and T helper 2 populations. J. Exp. Med. 182, 1357–1367.
Kamentski, L. A., Burger, D. E., Gersham, R. J., Kamentski, L. D., and Luther, E. (1997) Slide-based laser scanning cytometry. Acta. Cytol. 41, 123–143.
Andersson, J., Björk, L., Dinarello, C. A., and Andersson, U. (1992) Lipopolysacharide induces human interleukin-1 receptor antagonist and interleukin-1 in the same cell. Eur. J. Immunol. 22, 2617–2623.
Abrams, J. S., Roncarolo, H. G., Yssel, H., Andersson, U., Gleich, G. L., and Silver, J. E. (1992) Strategies of anti-cytokine monoclonal antibody development:Immonuassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 127, 5–24.
Ida, N. (1989) Establishment of strongly neautralizing mAb to human IL-6 and its epitope analysis. Biochem. Biophys. Res. Com. 165, 728–734.
Martich, G. D., Danner, R. L., Ceska, M., and Suffredini, A. F. (1991) Detection of IL-8 and TNF in normal humans after intravenous endotoxin. J. Exp. Med. 173, 1021–1024.
Lamche, H. R. and Adolf, G. R. (1990) Highly sensitive immunoassay for antibodies to human tumor necrosis factor and lymphotoxin. J. Immunol. Methods 131, 283–289.
Andersson, G., Ekre, H.-P., Alm, G., and Perlmann, P. (1989) Monoclonal antibody two-site ELISA for human IFN-?. J. Immunol. Methods 25, 89–96.
Gjörloff Wingren, A., Björkdahl, O., Labuda, T., Björk, L., Andersson, U., Gullberg, U., et al. (1996) Fusion of Signal Sequence to the Interleukin-1β gene directs the protein from cytoplasmatic accumulation to extracellular release. Cell. Immunol. 169, 226–237.
Willingham, M. C. and Pastan, I. (1985) An Atlas of Immunofluorescence in Cultured Cells. Academic Press Inc., London, UK, pp. 2–11.
Willingham, M. C. (1980) Electron microscopic immunocytochemical localization of intracellular antigens in cultured cells: the EGS and ferritin bridge procedures. Histochem. J. 12, 419–434.
Schneider, H., Vogt, A., and Bross, K. (1984) Identification of proliferating subpopulations in microcultures by surface marker and auto-radiography. Immunol. Commun. 13, 553–561.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Björk, L. (2003). Staining Protocol for Superantigen-Induced Cytokine Production Studied at the Single-Cell Level. In: Krakauer, T. (eds) Superantigen Protocols. Methods in Molecular Biology™, vol 214. Humana Press. https://doi.org/10.1385/1-59259-367-4:165
Download citation
DOI: https://doi.org/10.1385/1-59259-367-4:165
Publisher Name: Humana Press
Print ISBN: 978-0-89603-984-1
Online ISBN: 978-1-59259-367-5
eBook Packages: Springer Protocols