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Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers
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Published on: March 6, 2009

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CD4+ T Cell Epitope Identification from Complex Parasite Antigen Mixtures

Miguel Álvaro-Benito1, Friederike Ebner2, Miriam Bertazzon3

  • 1Laboratory of Protein Biochemistry, Institute of Chemistry and Biochemistry, Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin, Berlin, Germany. m.alvaro@fu-berlin.de.

Methods in Molecular Biology (Clifton, N.J.)
|May 31, 2023
PubMed

Antigen complexity represents a major challenge for scoring CD4+ T cell immunogenicity, a key hallmark of immunity and with great potential to improve vaccine development. In this chapter, we provide a comprehensive picture of a pipeline that can be applied to virtually any complex antigen to overcome different limitations. Antigens are characterized by Mass Spectrometry to determine the available protein sources and their abundances. A reconstituted in vitro antigen processing system is applied along with bioinformatics tools to prioritize the list of candidates. Finally, the immunogenicity of candidate peptides is validated ex vivo using PBMCs from HLA-typed individuals. This protocol compiles the essential information for executing the whole pipeline while focusing on the candidate epitope prioritizing scheme.

Keywords:
Antigen presentationAscaris spp. parasitesCD4+ T cell reactivityComplex antigen mixturesExcretory-secretory products (ES)MHC IIReconstitution in vitro antigen processing

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