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ELISPOT and intracellular cytokine staining: Novel assays for quantifying T cell responses in the chicken

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  • M. P. Ariaans
  • P. M. van de Haar
  • J. W. Lowenthal
  • W. van Eden
  • E. J. Hensen
  • L. Vervelde

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    Rights statement: Copyright 2008 Elsevier Ltd

    Final published version, 641 KB, PDF document

http://www.sciencedirect.com/science/article/pii/S0145305X08001122
Original languageEnglish
Pages (from-to)1398-1404
Number of pages7
JournalDevelopmental and Comparative Immunology
Volume32
Issue number11
DOIs
Publication statusPublished - 2008

Abstract

The measurement of T cell responses in chickens, not only for quantitative aspects but also for the qualitative nature of the responses, becomes increasingly important. However, there are very few assays available to measure T cell function. Therefore, we have developed enzyme-linked immunosorbent spot assay (ELISPOT) and an intracellular cytokine staining (ICCS) assay. ELISPOT assay for the detection of chicken interferon-gamma (ChIFN-gamma) production was set up and shown to be reproducible for both polyclonal and antigen-specific stimuli such as Newcastle disease virus (NDV). However, the ELISPOT assay lacks the ability to identify individual cytokine-producing cells. Separation of CD4(+) and CD8(+) T cell populations gave additional information, but appeared to have the disadvantage of a loss of cell interactions during stimulation. In a further refinement, individual cells were identifiable by ICCS, which gives the possibility to characterize for multiple characteristics, such as cytokine production and phenotype of the cell. Using ICCS, ChIFN-gamma production was evaluated. Although cells were detected at only low frequencies, polyclonal stimulation of peripheral, blood mononuclear cell (PBMC) or spleen cells resulted in a significant increase in ChIFN-gamma production by CD4(+) and CD8(+) cells. (C) 2008 Elsevier Ltd. All rights reserved.

    Research areas

  • LYMPHOCYTES, chicken, INTERFERON-GAMMA, STIMULATION, NEWCASTLE-DISEASE VIRUS, interferon-gamma, INFECTION, ELISPOT, T cell assay

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