This document summarizes research on the immune response to cytomegalovirus (CMV) and cancer-testis antigens (CTAgs). It discusses how CMV elicits a massive immune response, occupying up to 40% of CD8+ T cells. This response declines with immunosuppression. It also explores using CMV-specific T cells for immunotherapy after stem cell transplantation. The document then examines CTAgs, which are expressed in cancers and elicit anti-tumor immune responses, but challenges remain in understanding their efficacy.

1. Cytomegalovirus and cancer-specific immunity – lessons from across the spectrum of immunodominance Paul Moss School of Cancer Sciences

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. Led to collaboration on use of new reagents to study T cell immune responses Viral Antigen Fluorescence T Cell

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.

7. Herpes viruses Group of eight viruses HSV I & II, EBV, VZV, CMV, HHV-6 HHV-7, HHV-8

8. CMV infection is very common

10. CMV latency appears to be the result of chronic immune control Antibody NK cells T cells

11. T cell immunity in healthy donors

12. The CD8+ T cell response to CMV

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. How does CMV change the memory and naïve T cell count in healthy people ? T cells Naive Memory

15. The CD8+ memory T cell count is stable with age

16. CMV greatly increases the number of memory T cells

17. The CD8+ naïve T cell count declines with age

18. CMV reduces the naïve cell count at all ages

19. CMV seropositivity at age 50 years accelerates naïve T cell decline by 25 years

21. The clinical implications of CMV infection

22. Primary infection Time (years) Magnitude of T cell immunity

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. Immune senescence Time (years) Magnitude of T cell immunity

25. Does CMV infection contribute to immune senescence ?

26. Elderly individuals who are CMV seropositive die 4.5 years earlier than uninfected individuals

27. CMV-specific immunity in immunosuppressed patients

28. CMV was a major cause of death in the early days of stem cells transplantation CMV pneumonitis

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. Is it possible to transfer T cells into the patient to correct the early period of immunodeficiency ? CMV-specific T cells

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. Use tetramers to bind to CMV-specific T cells in the donor Stain T cells FACS analysis T Cell PE

33. T cells that bind tetramer can be selected with magnetic beads T Cell PE

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

40. Do we want to eradicate CMV ?

42. What is homo sapiens ? + + + 10 14 cells 10 15 cells a lot..

44. The immune response to cancer

48. MAGE-1 immunohistochemistry stain of human testis – Old LJ (2003)

49. testis ovary trophoblast Bladder cancer

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

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 

58. The CD8 response actually increases in the terminal stages of disease

59. MAGE – specific CD8 T cell clones can kill tumour when they are cloned in vitro (iii) (ii)

60. What about CD4 cells ?

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

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

65. CD4 CD8

66. CD4 immunity may determine the efficacy of the immune response CD4

67. The balance of inflammatory and regulatory CD4 immunity may be critical Th1 - IFN  Treg - suppression CD4 CD4

68. Th1 - IFN  Treg - suppression CD4 CD4

69. Th1 - IFN  Treg - suppression CD4 CD4

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