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Analysis of the Cellular Immune Responses to Vaccines

Nicholas Svitek1, Evans L N Taracha2, Rosemary Saya1

  • 1Animal and Human Health, International Livestock Research Institute, Nairobi, Kenya.

Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2022
PubMed

Insights

This study details methods for analyzing livestock immune responses using flow cytometry and ELISpot assays. These techniques evaluate cellular immunity 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 controlling intracellular pathogens and vaccine efficacy in livestock.
  • Assays like flow cytometry and ELISpot are vital for studying these responses in animal models.

Purpose of the Study:

  • To describe robust methods for evaluating cellular immunity in immunized livestock.
  • To present a comprehensive workflow for analyzing lymphocyte responses.

Main Methods:

  • Multiparametric flow cytometry using peptide-MHC class I tetramers and activation markers.
  • Interferon-gamma (IFN-γ) enzyme-linked immunospot (ELISpot) assays.
  • Chromium (51Cr) release assays for cellular cytotoxicity and a novel TCR avidity assay.

Main Results:

  • Detailed protocols for staining and analyzing bovine lymphocytes.
  • Demonstration of peptide-MHC tetramer staining for antigen-specific T cell identification.
  • Successful application of ELISpot and cytotoxicity assays for immune response assessment.

Conclusions:

  • Established methods enable detailed characterization of cellular immunity in livestock.
  • These assays are essential for vaccine development and understanding disease resistance in animals.
  • The described techniques facilitate precise evaluation of T cell responses and avidity.

Keywords:
CTL epitopeCytotoxicity assayELISpotFlow cytometryNetMHCpanPeptide-MHC class I tetramersTCR avidity

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