ELISPOT and intracellular cytokine staining: Novel assays for quantifying T cell responses in the chicken
| Autor: | W. van Eden, E. J. Hensen, John W. Lowenthal, Mark P. Ariaans, P.M. van de Haar, Lonneke Vervelde |
|---|---|
| Přispěvatelé: | Strategic Infection Biology, Dep Infectieziekten Immunologie |
| Rok vydání: | 2008 |
| Předmět: |
STIMULATION
T-Lymphocytes chicken medicine.medical_treatment T cell Immunology Cell Intracellular Space Mitosis Enzyme-Linked Immunosorbent Assay LYMPHOCYTES Peripheral blood mononuclear cell Interferon-gamma INFECTION medicine Animals Interferon gamma INTERFERON-GAMMA biology ELISPOT Vaccination Molecular biology T cell assay medicine.anatomical_structure Cytokine Polyclonal antibodies NEWCASTLE-DISEASE VIRUS biology.protein Chickens Spleen CD8 Developmental Biology medicine.drug |
| Zdroj: | Ariaans, M P, de Haar, P M V, Lowenthal, J W, van Eden, W, Hensen, E J & Vervelde, L 2008, ' ELISPOT and intracellular cytokine staining: Novel assays for quantifying T cell responses in the chicken ', Developmental and Comparative Immunology, vol. 32, no. 11, pp. 1398-1404 . https://doi.org/10.1016/j.dci.2008.05.007 |
| ISSN: | 0145-305X |
| DOI: | 10.1016/j.dci.2008.05.007 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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