| Literature DB >> 26458841 |
Nicholas Svitek1, Evans L N Taracha1, Rosemary Saya1, Elias Awino1, Vishvanath Nene1, Lucilla Steinaa2.
Abstract
Flow cytometry, enzyme-linked immunospot (ELISpot) and cellular cytotoxicity assays are powerful tools for studying the cellular immune response towards 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 towards a vaccine. Here, we describe staining of bovine lymphocytes with peptide (p)-MHC class I tetramers and antibodies specific towards cellular activation markers for evaluation by multiparametric flow cytometry, as well as interferon (IFN)-γ ELISpot and cytotoxicity using chromium ((51)Cr) release assays. A small component on the use of immunoinformatics for fine-tuning the identification of a minimal CTL epitope is included.Entities:
Keywords: CTL epitope; Cytotoxicity assay; ELISpot; Flow cytometry; NetMHCpan; Peptide-MHC class I tetramers
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Year: 2016 PMID： 26458841 DOI： 10.1007/978-1-4939-3008-1_16
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745