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Analysis of the Cellular Immune Responses to Vaccines

Nicholas Svitek1, Evans L N Taracha1, Rosemary Saya1

  • 1Vaccine Biosciences, International Livestock Research Institute, Att: Lucilla Steinaa, Vaccine Biosciences, 30709, 00100, Nairobi, Kenya.

Methods in Molecular Biology (Clifton, N.J.)
|October 14, 2015
PubMed

Insights

This study details methods for assessing livestock immune responses using flow cytometry, ELISpot, and cytotoxicity assays. These techniques evaluate lymphocyte reactivity to pathogens and vaccines, aiding in livestock health research.

Area of Science:

  • Veterinary Immunology
  • Cellular Immunology
  • Livestock Research

Background:

  • Cellular immune responses are crucial for livestock health, particularly against intracellular pathogens and vaccines.
  • Assessing these responses requires robust and specific analytical techniques.

Purpose of the Study:

  • To describe a comprehensive workflow for evaluating cellular immunity in livestock.
  • To detail methods for lymphocyte purification and analysis using advanced immunological assays.

Main Methods:

  • Ficoll-Paque density gradient centrifugation for lymphocyte isolation.
  • Multiparametric flow cytometry with peptide-MHC class I tetramers and activation markers.
  • Interferon-gamma (IFN-γ) Enzyme-Linked ImmunoSpot (ELISpot) assay.
  • Chromium ((51)Cr) release assay for cellular cytotoxicity.
  • Immunoinformatics for epitope identification.

Main Results:

  • Established protocols for multi-assay evaluation of bovine lymphocyte responses.
  • Demonstrated utility of peptide-MHC tetramers for identifying antigen-specific T cells.
  • Quantified cellular immune responses including IFN-γ production and cytotoxic activity.

Conclusions:

  • The described methods provide a powerful toolkit for dissecting cellular immunity in livestock.
  • These assays are essential for vaccine development and understanding disease resistance in agricultural animals.
  • Integration of immunoinformatics enhances epitope discovery for targeted immune interventions.

Keywords:
CTL epitopeCytotoxicity assayELISpotFlow cytometryNetMHCpanPeptide-MHC class I tetramers

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