Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
Cancer immunotherapy:  Finding allies among the ”allos” Can we induce clinical responses by identification of the right an...
Johanna Olweus ,  Radiumhospitalet ,  Oslo University Hospital ,  Oslo ,   Norway
Differences between the British and the Norwegians British architecture Norwegian architecture
 
 
BERGEN
 
The main function of the immune system is to protect against INFECTIONS It protects by recognizing and attacking  FOREIGN,...
High affinity T cells reactive with  self-antigens are negatively selected Negative selection of high affinity T cells rea...
Most tumor associated antigens (TAA) are self antigens expressed on a variety of normal cells Autologous immune responses ...
Survivin is widely expressed on normal healthy cells In Silico Transcriptomics online  Rolf Skotheim
Immunotherapy of cancer - what are the options?
Immunotherapy with antibodies - a success story Anti-CD20 mAbs in the treatment of leukemia/lymphoma <ul><li>Fastest growi...
Graft-versus-leukemia Modified from Bleakley et al, Nat Rev Cancer 2004 Can graft-versus-host reactive T cells be separate...
Can graft-versus-host reactive T cells be separated from graft-versus-leukemia reactive T cells? Transplanted donor T cell...
Models explaining the high frequencies of T cells recognizing allogeneic (foreign) HLA Allogeneic HLA: Peptide- dominant A...
Targeting foreign versus normal cell-type specific peptides Foreign  complex foreign self foreign self Vaccination targeti...
Protocol to generate allo-restricted T cells that kill specific cell types E Stronen et al, Scand J Immunol 2009; 69; 319-...
CD20/A2 pentamer+ T cells were generated from all A2 negative donors Donor T cells HLA-A*0201 neg Pentamer staining Cytoto...
CD20-targeted, allo-reactive T cells are specific for CD20  and HLA-A*0201 <ul><li>CTL lines stained negatively with >40 d...
Specificity: CD20-specific T cells are not activated by 4 different CD20neg target cells
CD20-specific T cells kill Chronic lymphocytic leukemia (CLL) cells CLL Cytotoxic T cell line I Abrahamsen et al, Leukemia...
CD20-specific T cells kill primary follicular lymphoma cells N=2 Ingerid Abrahamsen, Synneva Kjellevoll, submitted
The CD20-specific CTL line shows dependency on a large number of amino acids for peptide recognition CD20 WT peptide seque...
CD20-specific T cell clones are dependent on 6-9 amino acids for recognition of the CD20p I Abrahamsen et al, Leukemia, on...
Specificity of the CD20-specific T cells can be explained by a striking homology in their peptide-binding domains (CDR3b) ...
A foreign HLA-peptide complex can select highly peptide-specific T cells from donors with distinct MHC background <ul><li>...
Allogeneic HLA: Peptide- and HLA specific Autologous HLA : Peptide- and HLA specific Minor HAg Major HAg Foreign HLA-A*020...
T cell specific for cell-type restricted protein T cell receptor (TCR) + Patient T cells  ” Re-directed”  T cells killing ...
Predicting and screening candidate hematopoietic peptides Predicting peptides using computer algorithm from candidate prot...
Responses to 18/26  hematopoietic peptides * response at d26
Can you screen 10 peptides instead of 1 by pulsing the dendritic cells with 10 peptides separately? <ul><li>Conclusion:  <...
A polyclonal, multitargeted, high avidity attack!  CANCER CELL
<ul><li>The Olweus group </li></ul><ul><li>Ingerid Abrahamsen </li></ul><ul><li>Erlend Strønen </li></ul><ul><li>Lars-Egil...
Why does cancer develop in individuals with a well-functioning immune system? Cell infected with virus Cancer cell <ul><li...
 
Allo Foreign HLA-A2 Self HLA Peptide x Peptide y Auto OUR STRATEGY 1 Target  identification 2 Generation of T cells 4 TCR ...
Upcoming SlideShare
Loading in …5
×

ProImmune Antigen Characterization Summit Johanna Olweus

4,850 views

Published on

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"

Published in: Technology
  • Be the first to comment

ProImmune Antigen Characterization Summit Johanna Olweus

  1. 1. Cancer immunotherapy: Finding allies among the ”allos” Can we induce clinical responses by identification of the right antigens? <ul><li>Johanna Olweus </li></ul><ul><li>Dept.of Immunology, Institute for Cancer Research, </li></ul><ul><li>Radiumhospitalet, Oslo University Hospital and </li></ul><ul><li>the University of Oslo </li></ul>
  2. 2. Johanna Olweus , Radiumhospitalet , Oslo University Hospital , Oslo , Norway
  3. 3. Differences between the British and the Norwegians British architecture Norwegian architecture
  4. 6. BERGEN
  5. 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
  6. 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)
  7. 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?
  8. 11. Survivin is widely expressed on normal healthy cells In Silico Transcriptomics online Rolf Skotheim
  9. 12. Immunotherapy of cancer - what are the options?
  10. 13. Immunotherapy with antibodies - a success story Anti-CD20 mAbs in the treatment of leukemia/lymphoma <ul><li>Fastest growing class of drugs </li></ul><ul><li>Bind cell-type restricted proteins expressed on malignant and normal cells </li></ul>
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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%
  17. 20. CD20-targeted, allo-reactive T cells are specific for CD20 and HLA-A*0201 <ul><li>CTL lines stained negatively with >40 different HLA-A2 pentamers </li></ul>I Abrahamsen et al, Leukemia, online Sept 16, 2010 HEK293 cells % CD8 + T cells degranulating/ producing IFN-gamma
  18. 21. Specificity: CD20-specific T cells are not activated by 4 different CD20neg target cells
  19. 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
  20. 23. CD20-specific T cells kill primary follicular lymphoma cells N=2 Ingerid Abrahamsen, Synneva Kjellevoll, submitted
  21. 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
  22. 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
  23. 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
  24. 27. A foreign HLA-peptide complex can select highly peptide-specific T cells from donors with distinct MHC background <ul><li>The CD20-specific, allo-restricted T cells do not represent cross-reactive </li></ul><ul><li>T cells expanded by a pathogen presented on a shared, autologous HLA-ag </li></ul>
  25. 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
  26. 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
  27. 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
  28. 31. Responses to 18/26 hematopoietic peptides * response at d26
  29. 32. Can you screen 10 peptides instead of 1 by pulsing the dendritic cells with 10 peptides separately? <ul><li>Conclusion: </li></ul><ul><li>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 </li></ul>+ 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
  30. 33. A polyclonal, multitargeted, high avidity attack! CANCER CELL
  31. 34. <ul><li>The Olweus group </li></ul><ul><li>Ingerid Abrahamsen </li></ul><ul><li>Erlend Strønen </li></ul><ul><li>Lars-Egil Fallang </li></ul><ul><li>Jorunn Johansen </li></ul><ul><li>Synneva Kjellevoll </li></ul><ul><li>Mitsuko Komada </li></ul><ul><li>Shraddha Kumari </li></ul><ul><li>Sébastien Wälchli </li></ul><ul><li>Grant support: </li></ul><ul><li>Norwegian Research Council </li></ul><ul><li>Health Region South East </li></ul><ul><li>Oslo University Hospital </li></ul><ul><li>The Norwegian Cancer Society </li></ul><ul><li>Medinnova </li></ul><ul><li>Øyvind Andr é Foundation </li></ul><ul><li>National collaborators </li></ul><ul><li>Arne Kolstad </li></ul><ul><li>Geir Tjønnfjord </li></ul><ul><li>Gunnar Kvalheim </li></ul><ul><li>Inger Sandlie, IMMI, RH </li></ul><ul><li>Fridtjof Lund-Johansen, IMMI, RH </li></ul><ul><li>International </li></ul><ul><li>Ton Schumacher, The Netherlands Cancer Institute </li></ul><ul><li>Harlan Robins, Fred Hutch Cancer Research Center </li></ul><ul><li>Industry </li></ul><ul><li>Dynal (beads) </li></ul><ul><li>IBA (reversible tetramers for clin isolation) </li></ul>
  32. 35. Why does cancer develop in individuals with a well-functioning immune system? Cell infected with virus Cancer cell <ul><li>Cancer cells </li></ul><ul><li>may not be recognized as foreign </li></ul><ul><li>do not induce inflammation </li></ul><ul><li>secrete inhibitory factors </li></ul><ul><li>T CELLS ARE ANERGIZED OR </li></ul><ul><li>DELETED – LOW CLINICAL </li></ul><ul><li>RESPONSE RATE </li></ul><ul><li>FOR CANCER VACCINES </li></ul>Microbial antigens are FOREIGN Infection induces inflammation T CELLS CLEAR THE INFECTION
  33. 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

×