1. NK cell therapy after transplantation Miltenyi Satellite Symposium 1.4.2012

2. T-cell Immune Surveillance Pr ER Golgi

3. NK cell Immune Surveillance Pr ER Golgi NK + -

4. Killer Cell Immunoglobulin-like Receptors Inhibitory KIR allow NK cells to detect missing self (HLA).

5. Missing self in stem cell transplantation NK + - L L Leukemia patient Cw1/Cw4 Donor Cw1/Cw2

6. Stern, BMT, 2009 KIR mismatching in haploidentical HSCT

7. How to increase NK cell number and function Increase number of NK cells administered with graft  CD3/CD19 depletion vs. CD34 selection Adoptive transfer of NK cells  Ex vivo expanded versus in vivo expansion Enhancement of NK cell produced by the graft  Blocking of inhibitory receptors

8. Passweg Leukemia 2004 NK DLI to consolidate engraftment N=5 Infusion of non-expanded NK cells after haplo-HSCT Indication= mixed chimerism/relapse Hints of effectiveness

9. NK DLI to consolidate engraftment N=3 Infusion of IL-2-expanded (five-fold) NK cells after haplo-HSCT + IL-2 sc Indication= pre-emptive All patients achieved CR, 1 relapse/2 TRM Koehl, BCMD 2004

10. Preemptive NK DLI in high risk malignancies N=14 Infusion of NK cells derived from CD34+ PBSC (median 9*10e6/kg) Indication= pre-emptive No acute toxicity Yoon, BMT 2010

11. Infusion of NK DLI for relapse after haplo-HSCT Relapse after Haplo-HSCT despite KIR2DL1 ligand mismatch Re-induction with Mitoxantrone, Ara-C, Fludarabine Infusion of purified NK cells followed by IL-2 s.c. daily Nguyen, Transfusion 2011

12. What happened in the last 5-10 years? Shift to NK therapy without preceding transplant Establishment of GMP compatible expansion protocols

13. NK cell infusion after chemotherapy Miller Blood 2005 N=19 Infusion of CD3 depleted PBMC to patients with high-risk AML, s.c. IL-2 Engraftment of NK cells, requires lymphodepletion Temporary complete remission in 5 patients

14. NK cell infusion after chemotherapy Infusion of KIR ligand mismatched NK cells after Cy-Flu chemotherapy 10 pediatric patients with AML in CR1 Transient engraftment of donor NK cells, 100% PFS @ 2 years Rubnitz JCO 2010

15. Basel approach Siegler Cytotherapy 2010 OKT3, IL-2, irradiated autologous feeder cells

16. Expansion with modified K562 Denman Plos One 2012

17. Studies currently running or recently terminated More than half of all studies employ NK cells in the transplant setting

18. Conclusions Preparation of NK cell products technically feasible and safe for patients Exciting data coming in recent years from non-transplant settings Various competing approaches to produce expanded/activated NK cell products currently under evaluation Many studies currently evaluation NK cell therapy after transplantation

19. Thank you!