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Detection of Avian Antigen-Specific T Cells Induced by Viral Vaccines

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Vaccine Design

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1404))

Abstract

Live attenuated viral vaccines are widely used in commercial poultry production, but the development of new effective inactivated/subunit vaccines is needed. Studies of avian antigen-specific T cells are primarily based on analyses ex vivo after activating the cells with recall antigen. There is a particular interest in developing robust high-throughput assays as chicken vaccine trials usually comprise many individuals. In many respects, the avian immune system differs from the mammalian, and T cell assessment protocols must be adjusted accordingly to account for, e.g., differences in leukocyte subsets.

The carboxyfluorescein succinimidyl ester (CFSE) method described in this chapter has been adapted to chicken cells. In this test, cells of interest are stained with CFSE. The succinimidyl ester group covalently binds to cellular amines forming fluorescent conjugates that are retained in the cells even throughout division. This leads to daughter cells containing half the fluorescence of their parents. When lymphocytes are loaded with CFSE prior to ex vivo stimulation with specific antigen, the measurement of serial halving of its fluorescence by flow cytometry identifies the cells responding to the stimulation. This method has been successfully applied to studies of chicken antigen-specific T cells.

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References

  1. Davison TF (2003) The immunologists’ debt to the chicken. Br Poultry Sci 44:6–21

    Article  CAS  Google Scholar 

  2. Kaiser P (2012) The long view: a bright past, a brighter future? Forty years of chicken immunology pre- and post-genome. Avian Pathol 41:511–518

    Article  CAS  Google Scholar 

  3. Witter RL (2001) Marek’s disease vaccines – past, present and future – [chicken vs. virus – a battle of the centuries]. The Bart Rispens Memorial Lecture. Current Progress on Marek’s Disease Research 1–9

    Google Scholar 

  4. Liu G, Wang Q, Liu N, Xiao Y, Tong T, Liu S, Wu D (2012) Infectious bronchitis virus nucleoprotein specific CTL response is generated prior to serum IgG. Vet Immunol Immunopathol 148:353–358

    Article  CAS  Google Scholar 

  5. Niemiec PK, Read LR, Sharif S (2006) Synthesis of chicken major histocompatibility complex class II oligomers using a baculovirus expression system. Protein Expr Purif 46:390–400

    Article  CAS  Google Scholar 

  6. Kaiser P (2010) Advances in avian immunology – prospects for disease control: a review. Avian Pathol 39:309–324

    Article  CAS  Google Scholar 

  7. Kaiser P, Wu Z, Rothwell L, Fife M et al (2009) Prospects for understanding immune-endocrine interactions in the chicken. Gen Comp Endocrinol 163:83–91

    Article  CAS  Google Scholar 

  8. St Paul M, Paolucci S, Barjesteh N, Wood RD, Schat KA, Sharif S (2012) Characterization of chicken thrombocyte responses to Toll-like receptor ligands. PLoS One 7(8):e43381

    Article  CAS  Google Scholar 

  9. St Paul M, Paolucci S, Barjesteh N, Wood RD, Sharif S (2013) Chicken erythrocytes respond to Toll-like receptor ligands by up-regulating cytokine transcripts. Res Vet Sci 95:87–91

    Article  CAS  Google Scholar 

  10. Chang CF, Hamilton PB (1979) Thrombocyte as the primary circulating phagocyte in chickens. J Reticuloendoth Soc 25:585–590

    CAS  Google Scholar 

  11. Moser B, Eberl M (2007) gammadelta T cells: novel initiators of adaptive immunity. Immunol Rev 215:89–102

    Article  CAS  Google Scholar 

  12. Dalgaard TS, Norup LR, Rubbenstroth D, Wattrang E, Juul-Madsen HR (2010) Flow cytometric assessment of antigen-specific proliferation in peripheral chicken T cells by CFSE dilution. Vet Immunol Immunopathol 138:85–94

    Article  CAS  Google Scholar 

  13. Thiel A, Scheffold A, Radbruch A (2004) Antigen-specific cytometry – new tools arrived! Clin Immunol 111:155–161

    Article  CAS  Google Scholar 

  14. Picker LJ, Singh MK, Zdraveski Z, Treer JR et al (1995) Direct demonstration of cytokine synthesis heterogeneity among human memory/effector T cells by flow cytometry. Blood 86:1408–1419

    Article  CAS  Google Scholar 

  15. Suni MA, Picker LJ, Maino VC (1998) Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometry. J Immunol Methods 212:89–98

    Article  CAS  Google Scholar 

  16. Ariaans MP, van de Haar PM, Lowenthal JW, van Eden W et al (2008) ELISPOT and intracellular cytokine staining: novel assays for quantifying T cell responses in the chicken. Dev Comp Immunol 32:1398–1404

    Article  CAS  Google Scholar 

  17. Ruiz-Hernandez R, Peroval M, Boyd A, Balkissoon D et al (2014) An infected chicken kidney cell co-culture ELISpot for enhanced detection of T cell responses to avian influenza and vaccination. J Immunol Methods 416:40–48

    Article  Google Scholar 

  18. Dalgaard TS, Norup LR, Pedersen AR, Handberg KJ et al (2010) Flow cytometric assessment of chicken T cell-mediated immune responses after Newcastle disease virus vaccination and challenge. Vaccine 28:4506–4514

    Article  CAS  Google Scholar 

  19. Hou Y, Guo Y, Wu C, Shen N, Jiang Y, Wang J (2012) Prediction and identification of T cell epitopes in the H5N1 influenza virus nucleoprotein in chicken. PLoS One 7(6):e39344

    Article  CAS  Google Scholar 

  20. Norup LR, Dalgaard TS, Pedersen AR, Juul-Madsen HR (2011) Assessment of Newcastle disease-specific T cell proliferation in different inbred MHC chicken lines. Scand J Immunol 74:23–30

    Article  CAS  Google Scholar 

  21. Haghighi HR, Read LR, Haeryfar SM, Behboudi S, Sharif S (2009) Identification of a dual-specific T cell epitope of the hemagglutinin antigen of an h5 avian influenza virus in chickens. PLoS One 4(11):e7772

    Article  Google Scholar 

  22. Reemers SS, van Haarlem DA, Sijts AJ, Vervelde L, Jansen CA (2012) Identification of novel avian influenza virus derived CD8+ T-cell epitopes. PLoS One 7(2):e31953

    Article  CAS  Google Scholar 

  23. Ariaans MP, van de Haar PM, Hensen EJ, Vervelde L (2009) Infectious bronchitis virus induces acute interferon-gamma production through polyclonal stimulation of chicken leukocytes. Virology 385:68–73

    Article  CAS  Google Scholar 

  24. Miller MM, Bacon LD, Hala K, Hunt HD et al (2004) 2004 Nomenclature for the chicken major histocompatibility (B and Y) complex. Immunogenetics 56:261–279

    CAS  PubMed  Google Scholar 

  25. Barta O, Barta V, Pierson FW (1992) Optimum conditions for the chicken lymphocyte transformation test. Avian Dis 36:945–955

    Article  CAS  Google Scholar 

  26. Schaefer AE, Scafuri AR, Fredericksen TL, Gilmour DG (1985) Strong suppression by monocytes of T cell mitogenesis in chicken peripheral blood leukocytes. J Immunol 135:1652–1660

    CAS  PubMed  Google Scholar 

  27. Maecker HT, Ghanekar SA, Suni MA, He XS et al (2001) Factors affecting the efficiency of CD8+ T cell cross-priming with exogenous antigens. J Immunol 166:7268–7275

    Article  CAS  Google Scholar 

  28. Lavoie ET, Grasman KA (2005) Isolation, cryopreservation, and mitogenesis of peripheral blood lymphocytes from chickens (Gallus domesticus) and wild herring gulls (Larus argentatus). Arch Environ Contam Toxicol 48:552–558

    Article  CAS  Google Scholar 

  29. Brenchley JM, Douek DC (2004) Flow cytometric analysis of human antigen-specific T-cell proliferation. Methods Cell Biology 75:481–496

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by grants from the Danish Research Council (274-06-0258), the Danish Poultry Council, and Aarhus University.

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Authors and Affiliations

  1. Department of Animal Science, Aarhus University, Tjele, Denmark

    Tina Sørensen Dalgaard & Helle Risdahl Juul-Madsen

  2. Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark

    Liselotte Rothmann Norup

Authors
  1. Tina Sørensen Dalgaard
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  2. Liselotte Rothmann Norup
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  3. Helle Risdahl Juul-Madsen
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Corresponding author

Correspondence to Tina Sørensen Dalgaard .

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Editors and Affiliations

  1. Lankenau Institute for Medical Research, Wynnewood, PA, USA

    Sunil Thomas

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Dalgaard, T.S., Norup, L.R., Juul-Madsen, H.R. (2016). Detection of Avian Antigen-Specific T Cells Induced by Viral Vaccines. In: Thomas, S. (eds) Vaccine Design. Methods in Molecular Biology, vol 1404. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-3389-1_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3389-1_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-3388-4

  • Online ISBN: 978-1-4939-3389-1

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