Analysis of the cellular immune responses to vaccines
Flow cytometry, enzyme-linked immunospot (ELISpot), and cellular cytotoxicity assays are powerful tools for studying the cellular immune response toward intracellular pathogens and vaccines in livestock species. Lymphocytes from immunized animals can be purified using Ficoll-Paque density gradient c...
| Autores principales: | , , , , , |
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| Formato: | Capítulo de libro |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2022
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/118007 |
| _version_ | 1855525864048951296 |
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| author | Svitek, Nicholas Taracha, E.L.N. Saya, Rosemary Awino, Elias Nene, Vishvanath M. Steinaa, Lucilla |
| author_browse | Awino, Elias Nene, Vishvanath M. Saya, Rosemary Steinaa, Lucilla Svitek, Nicholas Taracha, E.L.N. |
| author_facet | Svitek, Nicholas Taracha, E.L.N. Saya, Rosemary Awino, Elias Nene, Vishvanath M. Steinaa, Lucilla |
| author_sort | Svitek, Nicholas |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Flow cytometry, enzyme-linked immunospot (ELISpot), and cellular cytotoxicity assays are powerful tools for studying the cellular immune response toward intracellular pathogens and vaccines in livestock species. Lymphocytes from immunized animals can be purified using Ficoll-Paque density gradient centrifugation and evaluated for their antigen specificity or reactivity toward a vaccine. Here, we describe staining of bovine lymphocytes with peptide (p)-MHC class I tetramers and antibodies specific toward cellular activation markers for evaluation by multiparametric flow cytometry, as well as interferon (IFN)-γ ELISpot and cytotoxicity using chromium (51Cr) release assays. A small component on the use of immunoinformatics for fine-tuning the identification of a minimal CTL epitope is included, and a newly developed and simple assay to measure TCR avidity. |
| format | Book Chapter |
| id | CGSpace118007 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | CGSpace1180072024-01-17T12:58:34Z Analysis of the cellular immune responses to vaccines Svitek, Nicholas Taracha, E.L.N. Saya, Rosemary Awino, Elias Nene, Vishvanath M. Steinaa, Lucilla vaccines immunology research Flow cytometry, enzyme-linked immunospot (ELISpot), and cellular cytotoxicity assays are powerful tools for studying the cellular immune response toward intracellular pathogens and vaccines in livestock species. Lymphocytes from immunized animals can be purified using Ficoll-Paque density gradient centrifugation and evaluated for their antigen specificity or reactivity toward a vaccine. Here, we describe staining of bovine lymphocytes with peptide (p)-MHC class I tetramers and antibodies specific toward cellular activation markers for evaluation by multiparametric flow cytometry, as well as interferon (IFN)-γ ELISpot and cytotoxicity using chromium (51Cr) release assays. A small component on the use of immunoinformatics for fine-tuning the identification of a minimal CTL epitope is included, and a newly developed and simple assay to measure TCR avidity. 2022 2022-02-08T13:12:28Z 2022-02-08T13:12:28Z Book Chapter https://hdl.handle.net/10568/118007 en Limited Access Springer Svitek, N., Taracha, E.L.N., Saya, R., Awino, E., Nene, V. and Steinaa, L. 2022. Analysis of the cellular immune responses to vaccines. IN: Brun, A. (ed), Vaccine Technologies for Veterinary Viral Diseases: Methods and Protocols, vol. 2465. New York, NY: Humana. pp. 283–301. |
| spellingShingle | vaccines immunology research Svitek, Nicholas Taracha, E.L.N. Saya, Rosemary Awino, Elias Nene, Vishvanath M. Steinaa, Lucilla Analysis of the cellular immune responses to vaccines |
| title | Analysis of the cellular immune responses to vaccines |
| title_full | Analysis of the cellular immune responses to vaccines |
| title_fullStr | Analysis of the cellular immune responses to vaccines |
| title_full_unstemmed | Analysis of the cellular immune responses to vaccines |
| title_short | Analysis of the cellular immune responses to vaccines |
| title_sort | analysis of the cellular immune responses to vaccines |
| topic | vaccines immunology research |
| url | https://hdl.handle.net/10568/118007 |
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