TY - CHAP
T1 - CD4+ T Cell Epitope Identification from Complex Parasite Antigen Mixtures
AU - Álvaro-Benito, Miguel
AU - Ebner, Friederike
AU - Bertazzon, Miriam
AU - Morrison, Eliot
N1 - Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - Antigen presentation
KW - Ascaris spp. parasites
KW - CD4+ T cell reactivity
KW - Complex antigen mixtures
KW - Excretory-secretory products (ES)
KW - MHC II
KW - Reconstitution in vitro antigen processing
UR - http://www.scopus.com/inward/record.url?scp=85160776111&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-3239-0_6
DO - 10.1007/978-1-0716-3239-0_6
M3 - Chapter
C2 - 37258908
AN - SCOPUS:85160776111
T3 - Methods in Molecular Biology
SP - 89
EP - 109
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -
