1. 1 Collaborations with Dr. PI Terasaki 1.Natural anti-human lymphocyte cytotoxic antibodies in mammalian sera (Ferrone S….., Terasaki PI: i) Transplantation. 1973;16:287-94. ii) Tissue Antigens. 1973;3:88-94). 2.Characterization of serum HLA antigens Ferrone S….., Terasaki PI: i) Tissue Antigens: 1975;5:41-7. ii) Immunol Commun. 1972;1:77-91). 3.HLA phenotyping of cultured B lymphoid cells (Ferrone S….., Terasaki PI: i) J Clin Invest. 1975;55:388-94. ii) Transplantation. 1976;22:61-8)

2. ( HLA class I antigens in tumor cells Clinical applications 1991-20111978-19902012- Hybridoma methodology Many HLA class I- specific mAbs Renaissance phase Indifferent phase Enthusiasm phase Anti-CTLA4 Anti-PD-1 Anti-PD-L1

3. Information in the literature 1.Defects in HLA class I antigens found in most if not all malignancies analyzed 2.Characterization of the multiple molecular mechanisms underlying HLA class I antigen defects in malignant cells 3.Assessment of the clinical significance of the HLA class I antigen defects found in malignant cells

4. ( HLA class I antigens in tumor cells Clinical applications 1991-20111978-19902012- Hybridoma methodology Many HLA class I- specific mAbs Renaissance phase Indifferent phase Enthusiasm phase Anti-CTLA4 Anti-PD-1 Anti-PD-L1

5. HLA Class I Antigen Processing in Tumors

6. 6 ERK1/2 activation in melanoma cells with a constitutively active BRAF-mutated

7. HLA Class I antigen up-regulation by BRAF-I and IFNα-2b on melanoma cells with an active BRAF-mutated SK-MEL-37 IFNα-2b BRAF-I HLA Class I mAb TP25.99HLA B/C mAb B1.23.2

8. 8 HLA Class I APM component up-regulation by EGFR-specific mAb cetuximab in the head and neck squamous cancer cell line JHU-029

9. 9 HLA-A2-MAGE-3 peptide complex up-regulation by EGFR-specific mAb cetuximab in the head and neck squamous cancer cell line JHU-029

10. 10 HLA Class I APM component up-regulation by docetaxel in human cancer cell lines

11. 11 Increased sensitivity to cognate CTL of human cancer cell lines treated with docetaxel

12. 12 HLA Class I APM component up-regulation by radiation in human cancer cell lines

13. 13 HLA class I antigen expression by malignant cells is modulated by signaling transduction pathways, chemotherapy and radiotherapy. These changes are associated with an increased T cell recognition and destruction of cancer cells. Conclusions

14. ( HLA class I antigens in tumor cells Clinical applications 1991-20111978-19902012- Hybridoma methodology Many HLA class I- specific mAbs Renaissance phase Indifferent phase Enthusiasm phase Anti-CTLA4 Anti-PD-1 Anti-PD-L1

15. Targeting Immunoregulatory Molecules in Cancer 15 Figure adapted from A. Ribas, NEJM, 2012

16. 16 Immunotherapy with check point molecule-specific mAb induces dramatic and long lasting responses, but only in up to 30% of patients. These findings emphasize the need to identify biomarkers to select patients who may benefit from this therapy HLA class I antigen expression by malignant cells may be a useful biomarker since defects in HLA class I antigen expression may protect tumor cells. Conclusions

17. 17

18. Soldano Ferrone, MD, PhD Departments of Surgey and Orthopedic Surgery Massachusetts General Hospital, Harvard Medical School, Boston, MA (USA) HLA class I antigen defects in malignant cells

19. 19 Acknowledgements Francesco Sabbatino, MD, PhD Cristina R. Ferrone, MD James Hodge, PhD (NCI, Bethesda, MD) Robert L. Ferris, MD, PhD (University of Pittsburgh, Pittsburgh, PA)

20. 20 Model of HLA Class I APM component up-regulation by docetaxel in human cancer cell lines

21. 21 HLA Class I antigen up-regulation by EGFR-specific mAb cetuximab on the head and neck squamous cancer cell line JHU-029