Detection of antigen-specific immunity (Cellular Immunology)

Sensitive detection of HLA-bound peptides on the tumor surface

T cell interactions with MHC presented peptides lie at the heart of any immune response. TIGL has developed and provides:

  • Detection of antigens, including CTAs, patient specific neoantigens, ERVs, and non-canonical peptides through optimized experimental workflows, mass spectrometry, and integrated data analysis, using clinical tissues samples
  • Serial HLA-peptide isolation methods enabling HLA-I and HLA-II isolation from the same sample
  • Mono-allelic systems to characterize the rules that govern HLA peptide processing and presentation
  • Proteogenomic approaches to generate patient-specific protein databases for antigen discovery
  • antigen-specific graph 1

    In our single-allele approach, B721.221 cells (~50 M) are transduced to express only one HLA allele. Immunopurified peptides are analyzed by LC-MS/MS and sequenced via an HLA-allele-specific database search (Abelin, Keskin et al Immunity 2017)

Immune-monitoring

The strength of TCR binding to cognate antigen (functional avidity) is strongly correlated with in vivo T cell effector function. TIGL has developed and provides:

  • ELISPOT assays and multicolor flow cytometry to investigate antigen-specific T cell responses
  • antigen-specific graph 2

    Example of IFN-γ ELISPOT results are shown for CD4+ T cells specific for mutated CDK13, JPH1, and ADM2d at week 16 after vaccination and after PD-1 blockade for patient 2. Complete response kinetics to mutated peptides are also shown (Ott and Hu et al Nature 2017).

TCR reconstruction

TIGL has developed and provides:

  • A novel TCR reconstruction pipeline that enables the testing of neoantigen-specific TCRs for functional avidity as an indicator of anti-tumor activity using ELISPOT and intracellular cytokine staining
  • antigen-specific graph 3

    A streamlined approach for identifying and reconstructing TCRs, and testing for specificity to candidate antigens (Hu, Anandappa et al Blood 2018).

Staff

Derin B. Keskin, PhD, Immunology Lead Scientist
Katarina Pinjusic, PhD, Postdoctoral Fellow
Allison Vanasse, Research Technician
Joseph Duke-Cohan, Research Technician, Bioinformatics
Melis Akinci, Research Technician
Marwa Belhaj, Research Technician