Gene and RNA Therapy Center (GRTC)

Peptide-based Immunotherapy

The overall goal of the Department of Peptide-based Immunotherapy is the development of novel immunotherapy concepts for malignant and infectious disease. Based on this, our research activity and expertise can be divided into four key areas:

• The direct mass spectrometry-based analysis of the immunopeptidome and the characterization of the antigen-specific T cell repertoire of malignant disease. With the combination of these unique techniques we provide a leading platform for the identification and characterization of tumor-associated antigens and for the evaluation of the dynamic of the tumor immunopeptidome and peptide-specific T cell response under the influence of drugs and stress conditions. 
• The characterization and optimization of antigen-specific T cell responses in infectious diseases. 
• The non-GMP and GMP production of peptides and peptide-based vaccines
• The translation of novel immunotherapeutic strategies from bench to bedside with a focus on peptide-based immunotherapies with currently four ongoing clinical trials evaluating own-developed peptide vaccines

List of GRT projects

Monitoring of T cell immunity for Gene and RNA therapies

Within these projects we apply our expertise in the characterization of antigenspeciif T cells to monitor the immunogenicity of gene and RNA therapeutics. For mRNA-based immunotherapies/vaccines the induction of T cell response is central for the clinical success of the compound (Maringer et al., Science Immunol 2022). For gene or protein-based therapeutics on the other hand immunogenicity i.e., specific T cell responses targeting the respective compound should be avoided. High-throughput T cell epitope screening and multiparametric analysis of antigen-specific T cell response is based on our unique prediction and selection workflow, integrating the algorithms SYFPEITHI and NetMHCpan.

This workflow enables the identification of dominant and highly immunogenic CD8⁺ and CD4⁺ T cell epitopes. Immunogenicity screening T cell epitopes is performed by IFN-γ enzyme-linked immunospot (ELISPOT) representing a well-established, highly sensitive method for the high throughput screening of T cell epitopes (Fig. 1A).

To even detect low frequent antigen-specific T cells, we established a protocol to expand antigen-specific T cells in vitro prior to ELISPOT analysis. Beside high-throughput identification of T cell epitopes this approach further enables the standardized large-scale analysis and quantification of patient cohorts for functional T cell responses to multiple HLA-matching antigens^38,44-46. For further multi-parametric characterization of these antigen-specific T cells we use a combinatorial approach of multi-parameter flow cytometry for T cell phenotype, T cell functionality and T cell exhaustion analysis (Fig. 1B) as well as two cytotoxicity assay systems (Vital assay, IncuCyte) evaluating specific lysis of peptide-loaded or peptide-presenting target cells by antigen-specific T cells (Fig. 1C).

GRT expertise

• Immunogenicity screening 
• Immunopeptidome analysis

Main GRT methods applied in the lab

• IFN-g ELISPOT
• Multi color flow cytometry
• Cytotoxicity assays
• Mass spectrometry based immunopeptidomics

List/description of clinical trials

Please refer to

Drug peptide laboratory | Universitätsklinikum Tübingen (uni-tuebingen.de)

Ongoing:

• B-pVAC-SARS-CoV-2; Prof. Dr. Juliane Walz; University Hospital Tübingen
• iVAC-L-CLL01; Prof. Dr. Helmut Salih; University Hospital Tübingen
• iVAC-XS15-CLL01; Prof. Dr. Juliane Walz; University Hospital Tübingen

 Completed:

• P-pVAC-SARS-CoV-2; Prof. Dr. Juliane Walz, University Hospital Tübingen (Heitmann JS et al, Nature)

 Upcoming:

• FusionVac_22, Prof. Dr. Juliane Walz, University Hospital Tübingen
• AML_VAC_XS1501, Prof. Dr. Juliane Walz, University Hospital Tübingen
• PerVision, PD. Dr. Martin Ebinger; University Hospital Tübingen

Ongoing and requested funding

Research | Universitätsklinikum Tübingen (uni-tuebingen.de)

• Jose Carreras Foundation
• Wilhelm Sander Foundation
• Bundesministerium für Bildung und Forschung (BMBF)
• Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg (MWK)
• Dieter Schwarz Stiftung
• Deutsche Krebshilfe
• Deutsches Konsortium für Translationale Krebsforschung (DKTK)
• Deutsche Forschungsgemeinschaft (DFG)
• Else Kröner Fresenius Stiftung
• iFIT Cluster of Excellence

Publications

Relevant links