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ProImmune Antigen Characterization Summit Johanna Olweus

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Johanna Olweus, Dept Immunology, Institute for Cancer Research, Radiumshospitalet, Oslo, Norway, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January …

Johanna Olweus, Dept Immunology, Institute for Cancer Research, Radiumshospitalet, Oslo, Norway, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Cancer immunotherapy: finding allies among the "allos"

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  • 1. Cancer immunotherapy: Finding allies among the ”allos” Can we induce clinical responses by identification of the right antigens?
    • Johanna Olweus
    • Dept.of Immunology, Institute for Cancer Research,
    • Radiumhospitalet, Oslo University Hospital and
    • the University of Oslo
  • 2. Johanna Olweus , Radiumhospitalet , Oslo University Hospital , Oslo , Norway
  • 3. Differences between the British and the Norwegians British architecture Norwegian architecture
  • 4.  
  • 5.  
  • 6. BERGEN
  • 7.  
  • 8. The main function of the immune system is to protect against INFECTIONS It protects by recognizing and attacking FOREIGN, DANGEROUS MATERIAL It does not attack self - TOLERANCE Important characteristics of the immune system
  • 9. High affinity T cells reactive with self-antigens are negatively selected Negative selection of high affinity T cells reactive with self-antigens Ed Palmer (May 2003)
  • 10. Most tumor associated antigens (TAA) are self antigens expressed on a variety of normal cells Autologous immune responses directed to TAA rarely give side effects, but also rarely give clinical responses High affinity TCRs yield more efficient anti-tumor responses Rosenberg S et al, J Immunol 2006 and Blood 2009 When TAA expressed on normal cells are targeted with high affinity T cells, toxicity and side effects occur Schendel et al, J Clin Invest 2010, Restifo et al PNAS 2008 Time to rethink cancer targets and identify high affinity T cells?
  • 11. Survivin is widely expressed on normal healthy cells In Silico Transcriptomics online Rolf Skotheim
  • 12. Immunotherapy of cancer - what are the options?
  • 13. Immunotherapy with antibodies - a success story Anti-CD20 mAbs in the treatment of leukemia/lymphoma
    • Fastest growing class of drugs
    • Bind cell-type restricted proteins expressed on malignant and normal cells
  • 14. Graft-versus-leukemia Modified from Bleakley et al, Nat Rev Cancer 2004 Can graft-versus-host reactive T cells be separated from graft-versus-leukemia reactive T cells? Graft-versus-host disease Patient cured Leukemia patient Allogeneic bone marrow/ stem cell donor
  • 15. Can graft-versus-host reactive T cells be separated from graft-versus-leukemia reactive T cells? Transplanted donor T cells Patient cells / organs GVL GVHD + -/+ - Goal: to generate allo-reactive T cells that specifically kill hematopoietic cells
  • 16. Models explaining the high frequencies of T cells recognizing allogeneic (foreign) HLA Allogeneic HLA: Peptide- dominant Allogeneic HLA: HLA dominant Autologous HLA : Peptide- and HLA specific Minor HAg Major HAg Major HAg Foreign Foreign Patient cell Donor T cell
  • 17. Targeting foreign versus normal cell-type specific peptides Foreign complex foreign self foreign self Vaccination targeting foreign peptide In vitro targeting Celltype specific peptides Peptides derived from cancer cells HLA antigen Peptide Danger signal
  • 18. Protocol to generate allo-restricted T cells that kill specific cell types E Stronen et al, Scand J Immunol 2009; 69; 319-28 CD20 derived peptide
  • 19. CD20/A2 pentamer+ T cells were generated from all A2 negative donors Donor T cells HLA-A*0201 neg Pentamer staining Cytotoxic T cells ” Patient APC” HLA-A*0201 Peptide specific for hematopoietic cells (CD20) HLA-A*0201neg. donors HLA-A*0201pos. donor I Abrahamsen et al, Leukemia, online Sept 16, 2010 CD8 CD20 pentamer 1.0% 2.8% 0.2% 0.1% 0.5% 0.6% CD8 Pentamer CD20 pentamer Control pentamer <0.01% <0.01%
  • 20. CD20-targeted, allo-reactive T cells are specific for CD20 and HLA-A*0201
    • CTL lines stained negatively with >40 different HLA-A2 pentamers
    I Abrahamsen et al, Leukemia, online Sept 16, 2010 HEK293 cells % CD8 + T cells degranulating/ producing IFN-gamma
  • 21. Specificity: CD20-specific T cells are not activated by 4 different CD20neg target cells
  • 22. CD20-specific T cells kill Chronic lymphocytic leukemia (CLL) cells CLL Cytotoxic T cell line I Abrahamsen et al, Leukemia, online Sept 16, 2010
  • 23. CD20-specific T cells kill primary follicular lymphoma cells N=2 Ingerid Abrahamsen, Synneva Kjellevoll, submitted
  • 24. The CD20-specific CTL line shows dependency on a large number of amino acids for peptide recognition CD20 WT peptide sequence: S L F L G I L S V Position: 1 2 3 4 5 6 7 8 9 I Abrahamsen et al, Leukemia, online Sept 16, 2010 Counts 100% 84.6% 155% 88.1% 35% 62.2% 42.9% 42.5% 102% 178% HLA-A*0201 multimer CD20 WT S188A L189A F190A L191A G192A I193A L194A S195A V196A 1 2 3 4 5 6 7 8 9
  • 25. CD20-specific T cell clones are dependent on 6-9 amino acids for recognition of the CD20p I Abrahamsen et al, Leukemia, online Sept 16, 2010 HLA-A*0201 multimer Counts CD20 WT S188A L189A F190A L191A G192A I193A L194A S195A V196A 1 2 3 4 6 7 8 9 5 1 2 3 4 5 6 7 8 9
  • 26. Specificity of the CD20-specific T cells can be explained by a striking homology in their peptide-binding domains (CDR3b) I Abrahamsen et al, Leukemia, online Sept 16, 2010
  • 27. A foreign HLA-peptide complex can select highly peptide-specific T cells from donors with distinct MHC background
    • The CD20-specific, allo-restricted T cells do not represent cross-reactive
    • T cells expanded by a pathogen presented on a shared, autologous HLA-ag
  • 28. Allogeneic HLA: Peptide- and HLA specific Autologous HLA : Peptide- and HLA specific Minor HAg Major HAg Foreign HLA-A*0201 Foreign HLA-A*0201/CD20 can select highly peptide-specific T cells from donors with distinct MHC background CD20-derived peptide
  • 29. T cell specific for cell-type restricted protein T cell receptor (TCR) + Patient T cells ” Re-directed” T cells killing leukemia cells HLA-A2 negative, T cell depleted bone marrow transplant to replace immune cells TCR transfer HLA-A2 positive leukemia/lymphoma patient Clone TCR peptide HLA-A2 Cell-type-specific T cell (from HLA-A2neg donor) leukemia Alternative 2 Alternative 1 Alt 1 Alt 2 Therapeutic strategies for adoptive T cell therapy
  • 30. Predicting and screening candidate hematopoietic peptides Predicting peptides using computer algorithm from candidate proteins Peptide synthesis and biochemical validation Pentamer synthesis Validation of epitopes in T cell cultures Søren Buus’ lab NetMHC ProImmmune Ltd. Olweus lab 150 peptides 70 peptide/MHC pentamers
  • 31. Responses to 18/26 hematopoietic peptides * response at d26
  • 32. Can you screen 10 peptides instead of 1 by pulsing the dendritic cells with 10 peptides separately?
    • Conclusion:
    • Similar immunogenicity when the peptide-pulsed DCs : T cell ratio is 1:100 as when it is 1: 10, provided that the overall DC : T cell ratio is 1:10
    + T cells + T cells + T cells DC:T cell ratio 1:10 peptideDC:T cell ratio 1:10 DC:T cell ratio 1:10 peptideDC:T cell ratio 1:100 DC:T cell ratio 1:100 peptideDC:T cell ratio 1:100
  • 33. A polyclonal, multitargeted, high avidity attack! CANCER CELL
  • 34.
    • The Olweus group
    • Ingerid Abrahamsen
    • Erlend Strønen
    • Lars-Egil Fallang
    • Jorunn Johansen
    • Synneva Kjellevoll
    • Mitsuko Komada
    • Shraddha Kumari
    • Sébastien Wälchli
    • Grant support:
    • Norwegian Research Council
    • Health Region South East
    • Oslo University Hospital
    • The Norwegian Cancer Society
    • Medinnova
    • Øyvind Andr é Foundation
    • National collaborators
    • Arne Kolstad
    • Geir Tjønnfjord
    • Gunnar Kvalheim
    • Inger Sandlie, IMMI, RH
    • Fridtjof Lund-Johansen, IMMI, RH
    • International
    • Ton Schumacher, The Netherlands Cancer Institute
    • Harlan Robins, Fred Hutch Cancer Research Center
    • Industry
    • Dynal (beads)
    • IBA (reversible tetramers for clin isolation)
  • 35. Why does cancer develop in individuals with a well-functioning immune system? Cell infected with virus Cancer cell
    • Cancer cells
    • may not be recognized as foreign
    • do not induce inflammation
    • secrete inhibitory factors
    • T CELLS ARE ANERGIZED OR
    • DELETED – LOW CLINICAL
    • RESPONSE RATE
    • FOR CANCER VACCINES
    Microbial antigens are FOREIGN Infection induces inflammation T CELLS CLEAR THE INFECTION
  • 36.  
  • 37. Allo Foreign HLA-A2 Self HLA Peptide x Peptide y Auto OUR STRATEGY 1 Target identification 2 Generation of T cells 4 TCR sequencing 3 a Epitope discovery 3 b Epitope selection 5 TCR expression/ modification 6 TCR selection 7 GMP development 8 Clinical trial 9 Immune monitoring