1. TUMOR IMMUNOLOGY ARPAD LANYI PhD

2. MORE THAN 100 TUMOR TYPES Most tumors develop later in life, mechanisms to control tumor development must exist. 14 million new cancer cases/year worldwide /50% mortality. 10 16 cell division in a lifetime. Each of us carries 60 or more germline mutations not present in parents. THE BIOLOGY OF TUMORS BRIEFLY

3. BENIGN OR MALIGNANT TUMORS

4. MULTIPLE MUTATIONS LEAD TO DEVELOPMENT OF TUMORS doi:10.1038/35101031 Tissue cells ACTIVATION OF ONCOGENES Mitogens Growth factor receptors Secondary messengers Transcriptional activators Cell cycle genes INACTIVATION OF TUMOR SUPPRESSOR GENES Growth inhibitors Cell cycle inhibitors Programmed cell death genes DNA repair enzymes Malignant cells

5. Oncogenesis OutcomeMechanismExample Growth promotion Overexpression of growth factor receptors (such as epidermal growth factor, or EGF) making cells more sensitive to growth stimuli c-erb-B2 Increased growth factor signal transduction by an oncogene that lacks the GTPase activity that limits GTP induction of cytoplasmic kinases that drive cell growth ras Overexpression of a gene product by stimulation from an oncogene (such as ras)c-sis Lack of normal gene regulation through translocation of a gene where it is controlled by surrounding genes to a place where it is no longer inhibited c-abl Binding of oncogene product to the nucleus with DNA transcriptional activation to promote entry into the cell cycle c-myc Loss of tumor suppressor gene function Lack of regulation of cell adhesion with loss of growth control through cell interactionAPC Loss of down-regulation of growth promoting signal transductionNF-1 Loss of regulation of cell cycle activation through sequestation of transcriptional factorsRb Loss of regulation of cell cycle activation through lack of inhibition of cell proliferation that allows DNA repair p53 Limitation of apoptosis Overexpression of genes, activated by translocation, prevents apoptosisbcl-2 ONCOGENESIS

6. ONCOGENEASSOCIATED TUMORS c-erb-B2Breast and ovarian carcinomas rasMany carcinomas and leukemias c-sisGliomas c-ablChronic myelogenous leukemia, acute lymphocytic leukemia c-mycLymphomas BRCA-1Breast and ovarian carcinomas APCColonic adenocarcinomas NF-1Neurofibromas and neurofibrosarcomas RbRetinoblastomas, osteosarcomas, small cell lung carcinomas p53Many carcinomas bcl-2Chronic lymphocytic leukemia, lymphomas VARIOUS ONCOGENES ARE ASSOCIATED WITH DEFINED TUMORS

7. TUMOR-ASSOCIATED VIRUSES

8. CHROMOSOMAL TRANSLOCATION IN BURKITT’S LYMPHOMA Uncontrolled proliferation due to the activation of c- myc oncogene. EBV induced tumor. 8148q-14q+ c-myc CH VH c-myc CH VH

9. THE HALLMARKS OF CANCER (OLD STYLE) Hanahan and Weinberg Cell, 2011 pp646- SO FAR INDEPENDENT OF THE IMMUNE SYSTEM

10. DO IMMUNOCYTES RECOGNIZE TUMORS? DOES THE IMMUNE SYSTEM CONTROL THE DEVELOPMENT OF TUMORS? (IMMUNOSURVEILLANCE) CAN THE IMMUNE SYSTEM BE MODIFIED TO ERADICATE TUMORS? SOME IMPORTANT QUESTIONS ARISE:

11. MHC-DEPENDENT REJECTION OF TUMORS

12. INFECTIVE FACIAL TUMOR IN THE INBREAD POPULATION OF THE TASMANIAN DEVIL About 2/3 of Hungary Population half million

13. TUMOR-SPECIFIC ANTIGENS AND TUMOR ASSOCIATED ANTIGENS

14. TUMOR-SPECIFIC ANTIGENS

15. CT ANTIGENS Cancer/testis antigens are expressed almost entirely by cancerous cells, showing little or no expression in healthy tissue, with the exception of normal testis, embryonic ovaries and placenta.

16. THE TUMOR-SPECIFIC IMMUNE RESPONSE

17. WHY ARE THEN TUMORS NOT ELIMINATED? Tumors are not highly immunogenic DOI: 10.1126/science.1203486 Limited cytotoxic T-cell activity: MHCI – mutation, β2m,TAP Apoptosis: soluble Fas TGF-β: inhibition of immune response, G1 arrest, loss of TGF-βR TOLEROGENICMICROENVIRONMENT

18. A tumor is a wound that never heals Hanahan and Weinberg Cell, 2011 pp646- THE HALLMARKS OF CANCER (ONE STEP FORWARD)

19. MECHANISMS OF TUMOR ESCAPE

20. MODULATION OF DC-FUNCTION BY THE TUMOR TSLP-INDUCED DCS RECRUIT TH2 AND TREG CELLS

21. MANIPULATION OF THE IMMUNE RESPONSE BY A TUMOR

22. MANY TUMORS LOOSE EXPRESSION OF HLA CLASS I PROTEINS Loss of HLA class I expression in prostate cancer Class I molecules are stained brown. The stain and HLA class I molecules are not seen on the tumor mass but are restricted to lymphocytes infiltrating the tumor and tissue stromal cells.

23. HUMAN EPITHELIAL TUMORS CAN INHIBIT THE RESPONSE OF LYMPHOCYTES EXPRESSING NKG2D

24. TUMOR IMMUNOTHERAPY

25. Sattwa S. Neelapu Mol. Oncol (2015 in press) MAIN STRATEGIES FOR CANCER IMMUNOTHERAPY

26. MONOCLONAL ANTIBODIES USED IN THE TREATMENT OF PATIENTS WITH CANCER ADCC---NK, macrophage, complement Immunotoxins

27. Antibodies bind to a cell-surface antigen of the tumor cells, for example CD20. The Fc regions of the antibodies engage FcγRIII on an NK cell, which then becomes activated to kill the tumor cell. MANY THERAPEUTIC ANTICANCER MONOCLONAL ANTIBODIES WORK BY DELIVERING THE TUMOR CELLS TO NK CELL- MEDIATED ADCC

28. ANTIBODIES ARE USED TO TARGET TOXINS TO TUMOR CELLS

29. CHECKPOINT THERAPIES Blocking negative receptors used by Tregs Blocking negative receptors used by Tregs Targeting stimulatory co-receptors with agonistic antibodies Targeting stimulatory co-receptors with agonistic antibodies

30. BLOCKING THE INHIBITORY EFFECTS OF CTLA4 WITH A HUMAN MONOCLONAL ANTIBODY (IPILIMUMAB ETC) Blocking the PD1/PDL1 interaction with a human monoclonal antibody also works. 2 year survival is over 24% but quite some complications (GI). Melanoma. Clinical trials: prostate cancer, lung carcinoma

31. Padmanee Sharma1,2 and James P. Allison SCIENCE 2015 Vol. 348 pp56 BLOCKING PD-1/PD-L1 INTERACTION WITH A HUMAN MONOCLONAL ANTIBODIES SIGNIFICANTLY IMPROVES THERAPY Works well for melanoma, but it is in trial for many less immunogenic tumors.

32. COMBINATIONAL THERAPIES MAY HELP WHEN TUMORS ARE NOT IMMUNOGENIC

33. AZ AKTÍV TUMOR-SPECIFIKUS IMMUNTERÁPIA LEHETŐSÉGEI A tumor antigének beviteli módja Tumor protein Tumor protein- derived peptide Anti-idiotipe Ab Vírus-tumor genome Plasmid DNAModified tumor cell Irradiated tumor cell Heat shock protein Modified DC Mocellin S et al. Lancet Oncology 2004 Tumor cell lysateLoaded DC IMMUNOTHERAPY BY VACCINATON

34. VACCINATION AGAINST HUMAN PAPILLOMA VIRUSES CAN PREVENT CERVICAL CANCER HPV is an oncogenic virus causing genital warts. 250000 women die of cervical cancer each year. Almost all cervical and ovarian cancers are HPV positive. Preventing chronic HPV infection should prevent cancer.

35. VACCINATION OF MELANOMA PATIENTS MAY CAUSE THEIR TUMOR TO REGRESS Rec. virusSynthetic peptide vaccine It is not yet clear when the vaccine would work. Spectrum from remission to no response.

36. ADOPTIVE TRANSFER OF DENDRITIC CELLS LOADED IN VITRO WITH TUMOR ANTIGENS

37. ADOPTIVE T-CELL TRANSFER

38. T-CELL RESPONSES TO TUMORS CAN BE IMPROVED WITH CHIMERIC ANTIGEN RECEPTORS (CARS) Low affinity of TCR (compared to virus specific T cells). MHC restriction prevents use in the entire population. Problem solved by Fv. Variable fragment of the heavy and light chains of a tumor-specific antibody made a single chain. Fusion of FV to an intracellular domain containing CD28, CD137 and zeta-chain sequences. Generates strong signal in the absence of costimulation.

39. B-CELL TUMORS CAN BE TARGETED BY CARS SPECIFIC TO CD19

40. TREATMENT OF B ‑ CELL TUMORS USING ANTI-CD19 CAR T CELLS

41. THANK YOU

42. WHY ARE THEN TUMORS NOT ELIMINATED? Tumours are not highly immunogenic DOI: 10.1126/science.1203486 Limited cytotoxic T-cell activity: MHCI – mutation, β2m,TAP Apoptosis: soluble Fas TGF-β: inhibition of immune respone, G1 arrest, loss of TGF-βR TOLEROGENICMICROENVIRONMENT