title: Analysis of the Cellular Immune Responses to Vaccines.
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.
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Svitek
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.
authors.forename: Evans L N
authors.initials: EL
authors.lastname: Taracha
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.
authors.forename: Rosemary
authors.initials: R
authors.lastname: Saya
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.
authors.forename: Elias
authors.initials: E
authors.lastname: Awino
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.
authors.forename: Vishvanath
authors.initials: V
authors.lastname: Nene
authors.affiliation: Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya. l.steinaa@cgiar.org.
authors.forename: Lucilla
authors.initials: L
authors.lastname: Steinaa
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D002417:Cattle; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D007111:Immunity, Cellular / Q000235:genetics / Q000276:immunology*; D058751:Livestock / Q000821:virology; D010455:Peptides / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014612:Vaccines / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D010455:Peptides; D014612:Vaccines
keywords: CTL epitope; Cytotoxicity assay; ELISpot; Flow cytometry; NetMHCpan; Peptide-MHC class I tetramers