Abstract
During systemic infection by Listeria monocytogenes the host develops a robust T cell-mediated immune response against the major immunodominant antigens of the pathogen. The enzyme-linked immuno-spot (ELISPOT) test is an accurate and reproducible means of measuring the extent of this T cell response. Here we describe a detailed ELISPOT protocol for measuring an epitope-specific CD8+ T cell-mediated immune response in mice vaccinated with low doses of L. monocytogenes. The basic approach can be easily adapted for the analysis of other vaccination regimes and target epitopes.
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References
Vijh S, Pamer EG (1997) Immunodominant and subdominant CTL responses to Listeria monocytogenes infection. J Immunol 158:3366–3371
Vazquez-Boland JA, Kuhn M, Berche P, Chakraborty T, Dominguez-Bernal G, Goebel W, Gonzalez-Zorn B, Wehland J, Kreft J (2001) Listeria pathogenesis and molecular virulence determinants. Clin Microbiol Rev 14:584–640
Pamer EG (2004) Immune responses to Listeria monocytogenes. Nat Rev Immunol 4:812–823
Miyahira Y, Murata K, Rodriguez D, Rodriguez JR, Esteban M, Rodrigues MM, Zavala F (1995) Quantification of antigen specific CD8+ T cells using an ELISPOT assay. J Immunol Methods 181:45–54
Bahey-El-Din M, Casey PG, Griffin BT, Gahan CG (2008) Lactococcus lactis-expressing listeriolysin O (LLO) provides protection and specific CD8(+) T cells against Listeria monocytogenes in the murine infection model. Vaccine 26:5304–5314
Bahey-El-Din M, Casey PG, Griffin BT, Gahan CG (2010) Expression of two Listeria monocytogenes antigens (P60 and LLO) in Lactococcus lactis and examination for use as live vaccine vectors. J Med Microbiol 59:904–912
McLaughlin HP, Bahey-El-Din M, Casey PG, Hill C, Gahan CG (2013) A mutant in the Listeria monocytogenes Fur-regulated virulence locus (frvA) induces cellular immunity and confers protection against listeriosis in mice. J Med Microbiol 62:185–190
Bahey-El-Din M, Casey PG, Griffin BT, Gahan CGM (2010) Efficacy of a Lactococcus lactis ΔpyrG vaccine delivery platform expressing chromosomally integrated hly from Listeria monocytogenes. Bioeng Bugs 1:66–74
Geginat G, Schenk S, Skoberne M, Goebel W, Hof H (2001) A novel approach of direct ex vivo epitope mapping identifies dominant and subdominant CD4 and CD8 T cell epitopes from Listeria monocytogenes. J Immunol 166:1877–1884
Stambas J, Pietersz G, McKenzie I, Nagabhushanam V, Cheers C (2002) Oxidised mannan-listeriolysin O conjugates induce Th1/Th2 cytokine responses after intranasal immunisation. Vaccine 20:1877–1886
Carvalho LH, Hafalla JC, Zavala F (2001) ELISPOT assay to measure antigen-specific murine CD8(+) T cell responses. J Immunol Methods 252:207–218
Acknowledgments
This work was supported by a grant from the Irish Health Research Board (grant number RP/2006/23) and by a grant to the Alimentary Pharmabiotic Centre from Science Foundation Ireland under the Centres for Science Engineering and Technology (CSET) programme (grant numbers O7/CE/B1368 and 12/RC/2273).
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Bahey-El-Din, M., Gahan, C.G.M. (2014). Vaccination Studies: Detection of a Listeria monocytogenes-Specific T Cell Immune Response Using the ELISPOT Technique. In: Jordan, K., Fox, E., Wagner, M. (eds) Listeria monocytogenes. Methods in Molecular Biology, vol 1157. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0703-8_22
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DOI: https://doi.org/10.1007/978-1-4939-0703-8_22
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-0703-8
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