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Cytomegalovirus and cancer-specific immunity –  lessons from across the spectrum of immunodominance Paul Moss School of Ca...
Overview <ul><li>Immunology of Cytomegalovirus </li></ul><ul><ul><li>Immune control in immune competent and immune suppres...
- HIV+ donors - Cloned HIV-specific CTL - Cloned TCR - cDNA from PBMC was probed with oligos for TCRs - Suggested effector...
Led to collaboration on use of new reagents to study T cell immune responses Viral Antigen Fluorescence T Cell
We could visualise HIV-specific T cells Altman, Moss et al Science. 1996 Oct 4;274(5284):94-6.Phenotypic analysis of antig...
Then used cytomegalovirus as a ‘control’ for HIV <ul><li>… but the immune response was bigger than HIV  </li></ul>
Herpes viruses Group of eight viruses HSV I & II,  EBV,  VZV,  CMV,  HHV-6 HHV-7, HHV-8
CMV infection is very common
CMV natural history <ul><li>CMV has evolved to co-exist with an  immunocompetent  host </li></ul><ul><li>CMV is poorly ada...
CMV latency appears to be the result of chronic immune control Antibody NK cells T cells
T cell immunity in healthy donors
The CD8+ T cell response to CMV
Age Percentage of CD8+ T cell response Khan et al , J.I. 2004 The CD8-specific T cell response to CMV increases with age D...
How does CMV change the  memory  and  naïve  T cell count in healthy people ? T cells Naive Memory
The CD8+ memory T cell count is stable with age
CMV greatly increases the number of memory T cells
The CD8+ naïve T cell count declines with age
CMV reduces the naïve cell count at all ages
CMV seropositivity at age 50 years accelerates naïve T cell decline by 25 years
The immune response to CMV is massive <ul><li>CD8:  2 - 40% of T cells  </li></ul><ul><li>CD4:  2 - 10% of T cells </li></...
The clinical implications of CMV infection
Primary infection Time (years) Magnitude of T cell immunity
Sometimes the initial (primary) infection with CMV can cause clinical problems “  I though it was the end of my tennis. Ev...
Immune senescence Time (years) Magnitude of T cell immunity
Does CMV infection contribute to immune senescence ?
Elderly individuals who are CMV seropositive die 4.5 years earlier than uninfected individuals
CMV-specific immunity in immunosuppressed patients
CMV was a major cause of death in the early days of stem cells transplantation CMV pneumonitis
Patients have very few CMV-specific T cells in the first three months following SCT Time post-SCT  Number of CMV-specific ...
Is it possible to transfer T cells into the patient to correct the early period of immunodeficiency ? CMV-specific T cells
BMT is the ideal setting for adoptive transfer of T cells D D P P Time Following SCT, the patient is immunologically toler...
Use tetramers to bind to CMV-specific T cells in the donor Stain T cells FACS analysis T Cell PE
T cells that bind tetramer can be selected with magnetic beads T Cell PE
Transferred T cells can  expand  and  clear  CMV viraemia ( Cobbold et al, JEM 2005) 2.10 6  CMV-specific T cells were giv...
 
 
 
 
What can we do to control CMV?  <ul><li>Anti-viral medication </li></ul><ul><ul><li>Will this reduce the CMV-specific immu...
Do we want to eradicate CMV ?
Could CMV have beneficial effects ? <ul><li>Allergy </li></ul><ul><li>Auto-immunity </li></ul><ul><li>Boost immunity to ot...
What is  homo sapiens ? + + + 10 14  cells 10 15  cells a lot..
Conclusions - Cytomegalovirus <ul><li>CMV is the most immunodominant pathogen for the human immune system </li></ul><ul><l...
The immune response to cancer
 
 
Cancer Testis Antigens (CTAgs) <ul><li>Proteins expressed by germline cells but  not  other normal tissues </li></ul><ul><...
MAGE-1 immunohistochemistry stain of human testis – Old LJ (2003)
testis ovary trophoblast Bladder cancer
Cancer-testis Antigens (CTAgs) in cancer <ul><li>•  Also expressed in many cancers </li></ul><ul><li>•  May reflect the ac...
Tumour regression following vaccination MAGE antigens in a melanoma patient  Evolution of skin metastases on the leg of pa...
MAGE proteins are expressed in myeloma   <ul><li>Van Baren 1999 </li></ul>
<ul><li>T cells purified from  myeloma patients  at  different stages of disease </li></ul><ul><li>cancer-testis antigen p...
PBMCs only PBMCs + RVRF PRE POST 0.05% 0.01% 0.00008   0.004   Example of CTAg-specific cell responses in myeloma CD8 IFN 
<ul><li>15 out of 37 myeloma patients showed a CTAg-specific T cell response which measured between 0.01 and 0.7% of the C...
Technical issues <ul><li>Used the cytokine secretion assay </li></ul><ul><li>Immune responses very difficult to detect wit...
Question concerning this T cell response <ul><li>Does it have any effective activity against the tumour  </li></ul><ul><li...
The CD8 response actually  increases  in the terminal stages of disease
MAGE   – specific CD8 T cell clones can kill tumour when they are cloned  in vitro (iii) (ii)
What about CD4 cells ?
Multiple myeloma has a pre-malignant phase <ul><li>Monoclonal gammopathy of uncertain significance (MGUS) </li></ul><ul><u...
Strong CD4+ responses can been seen in MGUS Pre-sort Post-sort DMSO  MAGE-A3 149-160  MAGE-A3 243-258   MAGE-A3 281-295 MA...
CD4 and CD8 responses show a different pattern in MGUS and myeloma Multiple Myeloma   MGUS Positive Negative Positive Nega...
Does CD4+ T cell immunity play a role in controlling the progression of MGUS ? <ul><li>T H 1 CD4+ T cell responses determi...
CD4 CD8
CD4 immunity may determine the efficacy of the immune response CD4
The balance of  inflammatory  and  regulatory  CD4 immunity may be critical Th1 - IFN  Treg - suppression CD4 CD4
Th1 - IFN  Treg - suppression CD4 CD4
Th1 - IFN  Treg - suppression CD4 CD4
Pattern of CTAg immunity <ul><li>Can they really protect against disease ? </li></ul><ul><li>Do healthy donors have subcli...
2000- renal transplant patient developed melanoma in kidney and breast 2000 - another transplant patient 2 developed melan...
Challenges <ul><li>Relatively low frequency response </li></ul><ul><li>Efficacy unclear </li></ul><ul><li>Potential role f...
Conclusions <ul><li>Cancer testis antigens may represent an important target for tumour-specific immune responses  </li></...
Acknowledgements <ul><li>Naeem Khan </li></ul><ul><li>Mark Cobbold </li></ul><ul><li>Batoul Pourghysari </li></ul><ul><li>...
 
 
 
 
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ProImmune Antigen Characterization Summit Paul Moss

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Paul Moss, School of Cancer Sciences, Birmingham UK, presents at the ProImmune Antigen Characterization and Biomarker Discovery Summit, January 2011.
Cytomegalovirus and Cancer-specific Immunity

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ProImmune Antigen Characterization Summit Paul Moss

  1. 1. Cytomegalovirus and cancer-specific immunity – lessons from across the spectrum of immunodominance Paul Moss School of Cancer Sciences
  2. 2. Overview <ul><li>Immunology of Cytomegalovirus </li></ul><ul><ul><li>Immune control in immune competent and immune suppressed donors </li></ul></ul><ul><ul><li>Immunotherapy </li></ul></ul><ul><li>Cancer Immunology </li></ul><ul><ul><li>Cancer testis antigens </li></ul></ul>
  3. 3. - HIV+ donors - Cloned HIV-specific CTL - Cloned TCR - cDNA from PBMC was probed with oligos for TCRs - Suggested effector frequency of 0.2 - 1%
  4. 4. Led to collaboration on use of new reagents to study T cell immune responses Viral Antigen Fluorescence T Cell
  5. 5. We could visualise HIV-specific T cells Altman, Moss et al Science. 1996 Oct 4;274(5284):94-6.Phenotypic analysis of antigen-specific T lymphocytes.
  6. 6. Then used cytomegalovirus as a ‘control’ for HIV <ul><li>… but the immune response was bigger than HIV </li></ul>
  7. 7. Herpes viruses Group of eight viruses HSV I & II, EBV, VZV, CMV, HHV-6 HHV-7, HHV-8
  8. 8. CMV infection is very common
  9. 9. CMV natural history <ul><li>CMV has evolved to co-exist with an immunocompetent host </li></ul><ul><li>CMV is poorly adapted to live with an immunosuppressed host </li></ul><ul><ul><li>transplant recipients </li></ul></ul><ul><ul><li>HIV infection </li></ul></ul>
  10. 10. CMV latency appears to be the result of chronic immune control Antibody NK cells T cells
  11. 11. T cell immunity in healthy donors
  12. 12. The CD8+ T cell response to CMV
  13. 13. Age Percentage of CD8+ T cell response Khan et al , J.I. 2004 The CD8-specific T cell response to CMV increases with age DE F P=0.0352 P0.0067 P=0.0159 P=0.07262 P=0.003
  14. 14. How does CMV change the memory and naïve T cell count in healthy people ? T cells Naive Memory
  15. 15. The CD8+ memory T cell count is stable with age
  16. 16. CMV greatly increases the number of memory T cells
  17. 17. The CD8+ naïve T cell count declines with age
  18. 18. CMV reduces the naïve cell count at all ages
  19. 19. CMV seropositivity at age 50 years accelerates naïve T cell decline by 25 years
  20. 20. The immune response to CMV is massive <ul><li>CD8: 2 - 40% of T cells </li></ul><ul><li>CD4: 2 - 10% of T cells </li></ul>Time (years) Magnitude of T cell immunity CMV
  21. 21. The clinical implications of CMV infection
  22. 22. Primary infection Time (years) Magnitude of T cell immunity
  23. 23. Sometimes the initial (primary) infection with CMV can cause clinical problems “ I though it was the end of my tennis. Even as a person I could feel myself changing. I just wanted to stay and home and not see anyone, not even my friends. But slowly I got better. I still have to be careful and I can’t train or work as hard as I once did” Justine Henin-Hardenne
  24. 24. Immune senescence Time (years) Magnitude of T cell immunity
  25. 25. Does CMV infection contribute to immune senescence ?
  26. 26. Elderly individuals who are CMV seropositive die 4.5 years earlier than uninfected individuals
  27. 27. CMV-specific immunity in immunosuppressed patients
  28. 28. CMV was a major cause of death in the early days of stem cells transplantation CMV pneumonitis
  29. 29. Patients have very few CMV-specific T cells in the first three months following SCT Time post-SCT Number of CMV-specific T cells
  30. 30. Is it possible to transfer T cells into the patient to correct the early period of immunodeficiency ? CMV-specific T cells
  31. 31. BMT is the ideal setting for adoptive transfer of T cells D D P P Time Following SCT, the patient is immunologically tolerant of cells taken from the donor
  32. 32. Use tetramers to bind to CMV-specific T cells in the donor Stain T cells FACS analysis T Cell PE
  33. 33. T cells that bind tetramer can be selected with magnetic beads T Cell PE
  34. 34. Transferred T cells can expand and clear CMV viraemia ( Cobbold et al, JEM 2005) 2.10 6 CMV-specific T cells were given 99.5% Pre-infusion blood sample 0% •  Prior to infusion the patient had CMV viraemia •  No CMV-specific T cells were present 0.5% 9 days later T cells were present and the patient became CMV negative Blood sample 9 days after infusion
  35. 39. What can we do to control CMV? <ul><li>Anti-viral medication </li></ul><ul><ul><li>Will this reduce the CMV-specific immune response ? </li></ul></ul><ul><ul><li>Starting trial of anti-viral drugs in elderly donors </li></ul></ul><ul><li>Vaccination </li></ul><ul><ul><li>Can we eradicate CMV ? </li></ul></ul>
  36. 40. Do we want to eradicate CMV ?
  37. 41. Could CMV have beneficial effects ? <ul><li>Allergy </li></ul><ul><li>Auto-immunity </li></ul><ul><li>Boost immunity to others pathogen </li></ul>
  38. 42. What is homo sapiens ? + + + 10 14 cells 10 15 cells a lot..
  39. 43. Conclusions - Cytomegalovirus <ul><li>CMV is the most immunodominant pathogen for the human immune system </li></ul><ul><li>Clinical features are subtle but potentially profound in immune competent donors </li></ul><ul><ul><li>These may potentially be modulated by anti-viral drugs </li></ul></ul><ul><li>Reactivation is a major concern in immune suppressed donors </li></ul><ul><ul><li>We are tackling this with cell therapy </li></ul></ul><ul><li>There are likely to be advantages for us in having CMV infection </li></ul><ul><ul><li>We need to use epidemiology to find our what these are </li></ul></ul>
  40. 44. The immune response to cancer
  41. 47. Cancer Testis Antigens (CTAgs) <ul><li>Proteins expressed by germline cells but not other normal tissues </li></ul><ul><li>There is a tesis-blood barrier so immune tolerance is not established against CTAgs </li></ul>
  42. 48. MAGE-1 immunohistochemistry stain of human testis – Old LJ (2003)
  43. 49. testis ovary trophoblast Bladder cancer
  44. 50. Cancer-testis Antigens (CTAgs) in cancer <ul><li>• Also expressed in many cancers </li></ul><ul><li>• May reflect the acquisition of a ‘germline-transcription profile’ </li></ul><ul><li>• This could be advantageous for survival, proliferation and invasion </li></ul><ul><li>This can lead to a cellular immune response that may be valuable against the tumour </li></ul>
  45. 51. Tumour regression following vaccination MAGE antigens in a melanoma patient Evolution of skin metastases on the leg of patient EB81 during treatment. van Baren N et al 2005 (A) (B) (C) (A) Before vaccination (B) After four vaccinations with ALVAC (after 3 months) (C) After 10 months
  46. 52. MAGE proteins are expressed in myeloma <ul><li>Van Baren 1999 </li></ul>
  47. 53. <ul><li>T cells purified from myeloma patients at different stages of disease </li></ul><ul><li>cancer-testis antigen peptides used to screen f o r tumour - specific T-cells using the cytokine secretion assay </li></ul>Study of the CD8+ T cell response to CTAg in Multiple Myeloma
  48. 54. PBMCs only PBMCs + RVRF PRE POST 0.05% 0.01% 0.00008 0.004 Example of CTAg-specific cell responses in myeloma CD8 IFN 
  49. 55. <ul><li>15 out of 37 myeloma patients showed a CTAg-specific T cell response which measured between 0.01 and 0.7% of the CD8+ T cell pool </li></ul>CTAg-specific CD8+ T cell immunity in myeloma patients Goodyear et al 2005, 2008
  50. 56. Technical issues <ul><li>Used the cytokine secretion assay </li></ul><ul><li>Immune responses very difficult to detect with HLA-peptide multimers </li></ul><ul><ul><li>Low frequency T cell response </li></ul></ul><ul><ul><li>Low affinity T cell binding </li></ul></ul>
  51. 57. Question concerning this T cell response <ul><li>Does it have any effective activity against the tumour </li></ul><ul><li>Is it just a useless immune response to proteins present within the tumour </li></ul><ul><li>How do the immune responses correlate with the clinical progression of the disease </li></ul>
  52. 58. The CD8 response actually increases in the terminal stages of disease
  53. 59. MAGE – specific CD8 T cell clones can kill tumour when they are cloned in vitro (iii) (ii)
  54. 60. What about CD4 cells ?
  55. 61. Multiple myeloma has a pre-malignant phase <ul><li>Monoclonal gammopathy of uncertain significance (MGUS) </li></ul><ul><ul><li>a small monoclonal paraprotein </li></ul></ul><ul><ul><li>no clinical damage </li></ul></ul><ul><ul><li>1% risk per year of developing myeloma </li></ul></ul>
  56. 62. Strong CD4+ responses can been seen in MGUS Pre-sort Post-sort DMSO MAGE-A3 149-160 MAGE-A3 243-258 MAGE-A3 281-295 MAGE-A1/A3 121-134 / MAGE-A3 191-205 SEB
  57. 63. CD4 and CD8 responses show a different pattern in MGUS and myeloma Multiple Myeloma MGUS Positive Negative Positive Negative CD4 1 29 4 26 CD8 13 42 1 weak 20
  58. 64. Does CD4+ T cell immunity play a role in controlling the progression of MGUS ? <ul><li>T H 1 CD4+ T cell responses determine clearance or progression of papilloma-virus infections </li></ul><ul><li>Indicates role for vaccination to boost CD4+ immune responses </li></ul>
  59. 65. CD4 CD8
  60. 66. CD4 immunity may determine the efficacy of the immune response CD4
  61. 67. The balance of inflammatory and regulatory CD4 immunity may be critical Th1 - IFN  Treg - suppression CD4 CD4
  62. 68. Th1 - IFN  Treg - suppression CD4 CD4
  63. 69. Th1 - IFN  Treg - suppression CD4 CD4
  64. 70. Pattern of CTAg immunity <ul><li>Can they really protect against disease ? </li></ul><ul><li>Do healthy donors have subclinical tumour cells ? </li></ul><ul><li>Can tumour cells remain latent for years ? </li></ul>
  65. 71. 2000- renal transplant patient developed melanoma in kidney and breast 2000 - another transplant patient 2 developed melanoma in kidney - Registry showed both patients had been transplanted from same patient in 1998 - This patient had received excision of 2.6mm melanoma in 1982 Tumour latency Mackie 2003
  66. 72. Challenges <ul><li>Relatively low frequency response </li></ul><ul><li>Efficacy unclear </li></ul><ul><li>Potential role for vaccination or adoptive transfer </li></ul>
  67. 73. Conclusions <ul><li>Cancer testis antigens may represent an important target for tumour-specific immune responses </li></ul><ul><li>CTAg-specific CD8+ T cells are detectable in patients after stem cell transplantation </li></ul><ul><li>CTAg expression can be upregulated by demethylation treatment in tumour cell lines </li></ul>
  68. 74. Acknowledgements <ul><li>Naeem Khan </li></ul><ul><li>Mark Cobbold </li></ul><ul><li>Batoul Pourghysari </li></ul><ul><li>Charles Craddock </li></ul><ul><li>Oliver Goodyear </li></ul><ul><li>Guy Pratt </li></ul>

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