Springer Protocols: Abstract: Measuring T-Cell Function in Animal Models of Tuberculosis by ELISPOT

Measuring T-Cell Function in Animal Models of Tuberculosis by ELISPOT

By: Vanja Lazarevic², Santosh Pawar², JoAnne Flynn²

Abstract

Full Text

Download PDF (197K)

Enzyme-linked immunospot (ELISPOT) was originally developed from an enzyme-linked immunosorbent assay (ELISA) to detect and measure the frequency of individual cells that produce cytokines in response to antigenic stimulation. ELISPOT assay is more sensitive than the enzyme-linked immunosorbent assay or intracellular cytokine staining. Increased sensitivity of ELISPOT is particularly advantageous when studying T-cell-mediated responses in Mycobacterium tuberculosis infection because antigen specific T-cells can occur at a low frequency in vivo. This method has been successfully used to analyze M. tuberculosis immune responses in humans in addition to murine and nonhuman primate models of tuberculosis.

Affiliation(s): (2) Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pittsburgh, PA

Book Title: Handbook of ELISPOT: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 302 | Pub. Date: Mar-15-2005 | Page Range: 179-189 | DOI: 10.1385/1-59259-903-6:179

Subject: Immunology

Key Words: Mycobacterium tuberculosis - IFN-γ - ELISPOT - bronchoalveolar lavage - lung - T-cells

References

Download References

1.	Chapman A. L., Munkanta M., Wilkinson K. A., Pathan A. A., Ewer K., Ayles H., et al. (2002). Rapid detection of active and latent tuberculosis infection in HIV-positive individuals by enumeration of Mycobacterium tuberculosis-specific T-cells. AIDS 16, 2285–2293.

2.	Ewer K., Deeks J., Alvarez L., Bryant G., Waller S., Andersen P., et al. (2003). Comparison of T-cell-based assay with tuberculin skin test for diagnosis of Mycobacterium tuberculosis infection in a school tuberculosis outbreak. Lancet 361, 1168–1173.

3.	Flynn J. L., Capuano S. V., Croix D., Pawar S., Myers A., Zinovik A., et al. (2003). Non-human primates: a model for tuberculosis research. Tuberculosis (Edinb) 83, 116–118.

4.	Hill P. C., Brookes R. H., Fox A., Fielding K., Jeffries D. J., Jackson-Sillah D., et al. (2004). Large-scale evaluation of enzyme-linked immunospot assay and skin test for diagnosis of Mycobacterium tuberculosis infection against a gradient of exposure in The Gambia. Clin Infect Dis 38, 966–973.

5.	Lalvani A., Pathan A. A., Durkan H., Wilkinson K. A., Whelan A., Deeks J. J., et al. (2001). Enhanced contact tracing and spatial tracking of Mycobacterium tuberculosis infection by enumeration of antigen-specific T-cells. Lancet 357, 2017–2021.

6.	Pathan A. A., Wilkinson K. A., Wilkinson R. J., Latif M., McShane H., Pasvol G., et al. (2000). High frequencies of circulating IFN-gamma-secreting CD8 cytotoxic T-cells specific for a novel MHC class I-restricted Mycobacterium tuberculosis epitope in M. tuberculosis-infected subjects without disease. Eur J Immunol 30, 2713–2721.

7.	Tsukaguchi K., Balaji K. N., and Boom W. H. (1995). CD4+ alpha beta T-cell and gamma delta T-cell responses to Mycobacterium tuberculosis. Similarities and differences in Ag recognition, cytotoxic effector function, and cytokine production. J Immunol 154, 1786–1796.

Comments (Loading...)

Post comment/View all comments