Methods: Patients with either ‘poor-risk’ or ‘very poor-risk’ leukemia were included in this phase I study. Either HLA matched siblings (MRD) or HLA matched (9 or 10/10) unrelated donors (MUD) were eligible. abT-cell reduction was performed by negative selection with anti-abTCR antibodies in combination with magnetic microbeads, using the automated CliniMACS device (Miltenyi Biotec, Bergisch Gladbach, Germany). The maximal contamination with abT-cells for all dose levels was 5x105/kg. The conditioning regimen consisted of: ATG (Genzyme®) 4 or 6 mg/m2 + fludarabine 120 mg/m2 + busilvex AUC=90 followed by αβT- cell depleted grafts from matched related or unrelated donors. No additional immune suppression was given after allo-SCT.
Results: Products for 15 patients have been successfully processed and used for αβT-cell depleted allo-SCT between 2013 and 2014. A ~4 log depletion of αβT-cells has been observed in the product with a recovery of ~75% of CD34+ cells. The combination of ATG/fludarabine/busilvex was well tolerated with a hematological recovery within 3 weeks. Primary engraftment (chimerism > 95%) was observed in all patients. Immune reconstitution primarily consisted of innate cells (NK cells and gd T cells) the first 6 months post transplantation. In addition, no increase in CMV or EBV reactivations has been observed so far under the profound “innate control”. Up to date, none of the patients developed aGVHD > grade II.
Conclusion: ATG Busulfan Fludarabine is a low toxicity platform for abTCR-depleted transplantations, resulting in a swift reconstitution of innate cells (NK cells and gd T cells) the first 6 months post transplantation. This transplantation strategy can serve as a tool for future immunological interventions such as a pre-emptive DLI or transfer of genetically modified T cells.