Abstract
The interferon-gamma (IFN-γ) release assay (IGRA) is an in vitro extension of the century-old in vivo tuberculin skin test, better known as the TST. Shortcomings to the TST are multifactorial and include limitations in sensitivity and specificity. IGRAs improve diagnostic specificity by using antigens not found in the Bacille Calmette-Guérin, a vaccine given in most countries. IGRAs capture the IFN-γ produced by T cells in response to antigen stimulation. The ELISPOT immediately captures IFN-γ produced directly from each cell, resulting in the generation of a cellular “footprint.” The dimensions and intensity of the generated footprint indicate the avidity of the secreting cell. We show a further improvement in IGRAs by addition of interleukin-7 (IL-7). IL-7 reduces T-cell apoptosis and stabilizes IFN-γ message. In addition to increasing the number of spots in the ELISPOT T-SPOT.TB platform, IL-7 increased IFN-γ production per cell as measured by an increase in spot size with no change in spot distribution.
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
Similar content being viewed by others
References
Lebedeva, Z. A. (1977) [Early Diagnosis of Tuberculosis (on the Centenary of the Birth of Charles Mantoux)]. Med Sestra 36, 51–52.
Lalvani, A., and Pareek, M. (2010) A 100 Year Update on Diagnosis of Tuberculosis Infection. Br Med Bull 93, 69–84.
Okada, K., Mao, T. E., Mori, T., Miura, T., Sugiyama, T., Yoshiyama, T., et al., (2008) Performance of an Interferon-Gamma Release Assay for Diagnosing Latent Tuberculosis Infection in Children. Epidemiol Infect 136, 1179–1187.
Diel, R., Loddenkemper, R., and Nienhaus, A. (2010) Evidence-Based Comparison of Commercial Interferon-γ Release Assays for Detecting Active TB. Chest 137, 952–968.
Lodha, R., and Kabra, S. K. (2004) Newer Diagnostic Modalities for Tuberculosis. Indian J Pediatr 71, 221–227.
Pai, M., Zwerling, A., and Menzies, D. (2008) Systematic Review: T-Cell--Based Assays for the Diagnosis of Latent Tuberculosis Infection: An Update. Ann Intern Med 149, 177–184.
Dheda, K., van, Z. S., Badri, M., and Pai, M. (2009) T-Cell Interferon-Gamma Release Assays for the Rapid Immunodiagnosis of Tuberculosis: Clinical Utility in High-Burden Vs. Low-Burden Settings. Curr Opin Pulm Med 15, 188–200.
Cattamanchi, A., Ssewenyana, I., Davis, J. L., Huang, L., Worodria, W., den Boon, S., et al., (2010) Role of Interferon-Gamma Release Assays in the Diagnosis of Pulmonary Tuberculosis in Patients with Advanced HIV Infection. BMC Infect Dis 10, 75–75.
Mazurek, G. H., Jereb, J., Lobue, P., Iademarco, M. F., Metchock, B., and Vernon, A. (2005) Guidelines for using the QuantiFERON-TB Gold Test for Detecting Mycobacterium Tuberculosis Infection, United States. MMWR Recomm Rep 54, 49–55.
Prabha, C., Karthic, S., Das, S. D., Swaminathan, S., Subramaniam, S., and Sukumar, B. (2005) Impact of Tuberculosis on Serum Leptin Levels in Patients with HIV Infection. Horm Res 63, 228–233.
Cellestis. QuantiFERON-TB Gold Package Insert. Cellestis.
Keeler, E., Perkins, M. D., Small, P., Hanson, C., Reed, S., Cunningham, J., et al., (2006) Reducing the Global Burden of Tuberculosis: The Contribution of Improved Diagnostics. Nature 444 Suppl 1, 49–57.
Pai, M., Minion, J., Sohn, H., Zwerling, A., and Perkins, M. D. (2009) Novel and Improved Technologies for Tuberculosis Diagnosis: Progress and Challenges. Clin Chest Med 30, 701.
Ewer, K., Millington, K. A., Deeks, J. J., Alvarez, L., Bryant, G., and Lalvani, A. (2006) Dynamic Antigen-Specific T-Cell Responses After Point-Source Exposure to Mycobacterium Tuberculosis. Am J Respir Crit Care Med 174, 831–839.
Klinman, D. (2008) ELISPOT Assay to Detect Cytokine-Secreting Murine and Human Cells. Curr Protoc Immunol Chapter 6, 6.19-6.19.
Kalyuzhny, A. E. (2005) Handbook of ELISPOT: Methods and Protocols, in Methods in molecular biology, 302 Totowa, N.J.: Humana Press, c2005.
Schlingmann, T. R., Shive, C. L., Targoni, O. S., Tary-Lehmann, M., and Lehmann, P. V. (2009) Increased Per Cell IFN-γ Productivity Indicates Recent in Vivo Activation of T Cells. Cell Immunol 258, 131–137.
CDC. (2000) Targeted Tuberculin Testing and Treatment of Latent Tuberculosis Treatment. CDC MMWR.
Pai, M., Riley, L. W., and Colford, John M, Jr. (2004) Interferon-Gamma Assays in the Immunodiagnosis of Tuberculosis: A Systematic Review. Lancet Infect Dis 4, 761–776.
Orlando, G., Merli, S., Cordier, L., Mazza, F., Casazza, G., Villa, A. M., et al., (2010) Interferon-Gamma Releasing Assay Versus Tuberculin Skin Testing for Latent Tuberculosis Infection in Targeted Screening Programs for High Risk Immigrants. Infection 38, 195–204.
Leyten, E. M. S., Arend, S. M., Prins, C., Cobelens, F. G. J., Ottenhoff, T. H. M., and van Dissel, J. T. (2007) Discrepancy between Mycobacterium Tuberculosis-Specific Gamma Interferon Release Assays using Short and Prolonged in Vitro Incubation. Clin Vaccine Immunol 14, 880–885.
Grimes, C. Z., Hwang, L., Williams, M. L., Austin, C. M., and Graviss, E. A. (2007) Tuberculosis Infection in Drug Users: Interferon-Gamma Release Assay Performance. Int J Tuberc Lung Dis 11, 1183–1189.
Helms, T., Boehm, B. O., Asaad, R. J., Trezza, R. P., Lehmann, P. V., and Tary-Lehmann, M. (2000) Direct Visualization of Cytokine-Producing Recall Antigen-Specific CD4 Memory T Cells in Healthy Individuals and HIV Patients. J Immunol 164, 3723–3732.
Marrack, P., and Kappler, J. (2004) Control of T Cell Viability. Annu Rev Immunol 22, 765–787.
Flynn, J. L., and Chan, J. (2001) Immunology of Tuberculosis. Annu Rev Immunol 19, 93.
Ott, P. A., Tary-Lehmann, M., and Lehmann, P. V. (2007) The Secretory IFN-γ Response of Single CD4 Memory Cells After Activation on Different Antigen Presenting Cell Types. Clinical Immunology 124, 267–276.
Rosenberg, S. A., Sportès, C., Ahmadzadeh, M., Fry, T. J., Ngo, L. T., Schwarz, S. L., et al., (2006) IL-7 Administration to Humans Leads to Expansion of CD8+ and CD4+ Cells but a Relative Decrease of CD4+ T-Regulatory Cells. J Immunother 29, 313–319.
Fry, T. J., and Mackall, C. L. (2005) The Many Faces of IL-7: From Lymphopoiesis to Peripheral T Cell Maintenance. J Immunol 174, 6571–6576.
Li, Masucci, Levitsky, and Levitsky, V. (2000) Effect of Interleukin-7 on the in Vitro Development and Maturation of Monocyte Derived Human Dendritic Cells. Scand. J Immunol 51, 361–371.
Carreno, B. M., Becker-Hapak, M., and Linette, G. P. (2009) CD40 Regulates Human Dendritic Cell-Derived IL-7 Production that, in Turn, Contributes to CD8(+) T-Cell Antigen-Specific Expansion. Immunol Cell Biol 87, 167–177.
Feske, M., Nudelman, R. J., Medina, M., Lew, J., Singh, M., Couturier, J., et al., (2008) Enhancement of Human Antigen-Specific Memory T-Cell Responses by Interleukin-7 may Improve Accuracy in Diagnosing Tuberculosis. Clin Vaccine Immunol 15, 1616–1622.
Lehmann, P. V. (2005) Image Analysis and Data Management of ELISPOT Assay Results. Methods Mol Biol 302, 117–132.
Hesse, M. D., Karulin, A. Y., Boehm, B. O., Lehmann, P. V., and Tary-Lehmann, M. (2001) A T Cell Clone’s Avidity is a Function of its Activation State. J Immunol 167, 1353–1361.
Slifka, M. K., and Rodriguez, F. (1999) Rapid on/off Cycling of Cytokine Production by Virus-Specific CD8+ T Cells. Nature 401, 76.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Feske, M.L., Medina, M., Graviss, E.A., Lewis, D.E. (2012). IL-7 Addition Increases Spot Size and Number as Measured by T-SPOT.TB ® . In: Kalyuzhny, A. (eds) Handbook of ELISPOT. Methods in Molecular Biology, vol 792. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-325-7_18
Download citation
DOI: https://doi.org/10.1007/978-1-61779-325-7_18
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-324-0
Online ISBN: 978-1-61779-325-7
eBook Packages: Springer Protocols