Abstract
The main characteristic of a superantigen (SAg) is that it activates specific subsets of T cells that are composed of particular T-cell receptor (TCR) variable region β chain gene products (TCRBV). SAgs are thought to form a bridge between the TCRBV region on the T cell and the MHC class II molecule on the antigen-presenting cell (APC). This bridging transduces a strong signal to the T cell through the TCR, resulting in T-cell activation, which is manifested by upregulation of cell-surface markers associated with activation, T-cell proliferation, and cytokine release. In this chapter we shall outline two methods for assessing specific T-cell activation based on the preceding criteria: 1) detection of markers of activation in individual TCRBV subsets early after SAg stimulation; and 2) analysis of cytokine release from TCRBV specific T-cell hybridomas in response to SAg presentation.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Herman, A., Kappler, J. W., Marrack, P., and Pullen, A. M. (1991) Super antigens: mechanism of T-cell stimulation and role in immune responses. Annu. Rev. Immunol. 9, 745–772.
Testi, R., Pulcinelli, F., Frati, L., Gazzaniga, P. P., and Santoni, A. (1990) CD69 is expressed on platelets and mediates platelet activa-tion and aggregation. J. Exp. Med. 172, 701–707.
Sutkowski, N., Palkama, T., Ciurli, C., Sekaly, R. P., Thorley-Lawson, D. A., and Huber, B. T. (1996) An Epstein-Barr virus-asso-ciated superantigen. J. Exp. Med. 184, 971–980.
Waller, E. K., Sen-Majumdar, A., Kamel, O. W., Hansteen, G. A., Schick, M. R., and Weissman, I. L. (1992) Human T-cell development in SCID-hu mice: staphylococcal enterotoxins induce specific clonal deletions, proliferation, and anergy. Blood 80, 3144–3156.
Baribaud, F., Wirth, S., Maillard, I., Valsesia, S., Acha-Orbea, H., and Diggelmann, H. (2001) Identification of key amino acids of the mouse mammary tumor virus superantigen involved in the specific interaction with T-cell receptor V(beta) domains. J. Virol. 75, 7453–7461.
Melchers, F., Potter, M., and Warner, N. L. (1978) Lymphocyte hybridomas. Second workshop on "e;functional properties of tumors of T and B lymphocytes."e; Preface. Curr. Top. Microbiol. Immunol. 81, IX–XXIII.
Kouttab, N., Tannir, N., Pathak, S., Berger, A., Sahasrabuddhe, C. G., and Maizel, A. L. (1985) Establishment of stable human T-T hybridomas. J. Immunol. Methods 77, 165–172.
Choi, Y. W., Herman, A., DiGiusto, D., Wade, T., Marrack, P., and Kappler, J. (1990) Residues of the variable region of the T-cell-receptor beta-chain that interact with S. aureus toxin superantigens. Nature 346, 471–473.
Kappler, J. W., Skidmore, B., White, J., and Marrack, P. (1981) Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition. J. Exp. Med. 153, 1198–1214.
Norcross, M. A., Bentley, D. M., Margulies, D. H., and Germain, R. N. (1984) Membrane Ia expression and antigen-presenting accessory cell function of L cells transfected with class II major histocompatibility complex genes. J. Exp. Med. 160, 1316–1337.
Mix, D. and Winslow, G. (1996) Proteolytic processing activates a viral superantigen. J. Exp. Med. 184, 1549–1554.
Conlon, P. J. (1983) A rapid biologic assay for the detection of interleukin 1. J. Immunol. 131, 1280–1282.
Hori, T., Uchiyama, T., Tsudo, M., Umadome, H., Ohno, H., Fukuhara, S., et al. (1987) Establishment of an interleukin 2-depen-dent human T cell line from a patient with T cell chronic lymphocytic leukemia who is not infected with human T cell leukemia/lymphoma virus. Blood 70, 1069–1072.
Beutner, U., Rudy, C., and Huber, B. T. (1992) Molecular characterization of Mls-1. Int. Rev. Immunol. 8, 279–288.
Sutkowski, N., Conrad, B., Thorley-Lawson, D. A., and Huber, B. T. (2001) Epstein-Barr virus transactivates the human edogenous retro virus HERV-K18 that encodes a superantigen. Immunity 15, 579–589.
Staufer, Y., Marguerat, S., Meylan, F., Ucla, C., Sutkowski, N., Huber, B. T., Pelet, T., and Conrad, B. (2001) Interferon-α induced endogenous superantigen: a model link between the environment and autoimmunity. Immunity 15, 591–601.
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
Sutkowski, N., Huber, B.T. (2003). Assessment of Specific T-Cell Activation by Superantigens. In: Krakauer, T. (eds) Superantigen Protocols. Methods in Molecular Biology™, vol 214. Humana Press. https://doi.org/10.1385/1-59259-367-4:185
Download citation
DOI: https://doi.org/10.1385/1-59259-367-4:185
Publisher Name: Humana Press
Print ISBN: 978-0-89603-984-1
Online ISBN: 978-1-59259-367-5
eBook Packages: Springer Protocols