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Background:Donor lymphocyte infusion (DLI) is commonly used to enhance T-cell mediated graft-versus-tumor (GVT) effect after allogeneic stem cell transplantation (allo-HSCT). It was showed that prophylactic DLI can produce promising outcome but is also associated with a high rate of graft-versus-host disease (GVHD), limiting its use as maintenance treatment. By contrast with T cells, there are clinical and biological evidences that NK cells can provide strong GVT effect without inducing GVHD, making post allo-HSCT NK cell-based immunotherapy appealing. Thus, we designed a phase 1 clinical trial evaluating the safety of donor-derived ex vivo IL-2 activated NK cells (DLI-NK) as maintenance immunotherapy after allo-HSCT (DLI-NK: NCT01853358).

Recent reports linking insertional activation of LMO2following gene therapy for X-linked severe combined immunodeficiency (X-SCID) have led to a re-evaluation of risks following gene therapy with retroviral vectors. In our analysis of 702 integration sites in rhesus macaques that underwent transplantation up to 7 years earlier with autologous CD34+cells transduced with amphotropic murine leukemia virus (MLV)-derived retroviral vectors containing marker genes, we detected insertion into one locus, the Mds1/Evi1region, a total of 14 times in 9 animals. Mds1/Evi1integrations were observed stably long term, primarily in myeloid cells. We hypothesize that this over-representation likely results from an impact on the self-renewal and engraftment potential of CD34+progenitor cells via insertional mutagenesis at this specific locus. There is no evidence of ongoing in vivo clonal expansion of the Mds1/Evi1populations, and all animals are hematologically normal without evidence for leukemia. Characterization of integration sites in this relevant preclinical model provides critical information for gene therapy risk assessment as well as identification of genes controlling hematopoiesis. (Blood. 2005;106:2530-2533)

Recent reports linking insertional activation of LMO2 following gene therapy for X-linked severe combined immunodeficiency (X-SCID) have led to a re-evaluation of risks following gene therapy with retroviral vectors. In our analysis of 702 integration sites in rhesus macaques that underwent transplantation up to 7 years earlier with autologous CD34+ cells transduced with amphotropic murine leukemia virus (MLV)-derived retroviral vectors containing marker genes, we detected insertion into one locus, the Mds1/Evi1 region, a total of 14 times in 9 animals. Mds1/Evi1 integrations were observed stably long term, primarily in myeloid cells. We hypothesize that this over-representation likely results from an impact on the self-renewal and engraftment potential of CD34+ progenitor cells via insertional mutagenesis at this specific locus. There is no evidence of ongoing in vivo clonal expansion of the Mds1/Evi1 populations, and all animals are hematologically normal without evidence for leukemia. Characterization of integration sites in this relevant preclinical model provides critical information for gene therapy risk assessment as well as identification of genes controlling hematopoiesis. (Blood. 2005;106:2530-2533)

Due to high frequency of side effects caused by high-dose total body irradiation (TBI) the nonmyeloablative regimen together with cytotoxic agents is currently used especially for elderly patients. However, immediate and long-term effects of low-dose irradiation used in allogeneic transplantation on stem cells is less well known. We have studied the effect of low-dose 3 Gy TBI on the number of hematopoietic stem cell (HSC) clones contributing simultaneously to granulocyte production in rhesus macaque. The number of clones after 3 Gy TBI decreased markedly by 2 to 3 weeks after 3 Gy TBI, followed by a period of clonal instability, and recovery to almost pre–3 Gy TBI clonal diversity. The clones accounting for this recovery contributed before 3 Gy TBI, suggesting the profound initial impact of TBI was on a pool of progenitor cells, whereas most of the more primitive HSCs remained unaffected and were able to again contribute to hematopoiesis after recovery. Clonal fluctuation may indirectly suggest the presence of short-term/long-term HSC populations in rhesus macaque bone marrow as reported in a mouse model. The results indicate that even low-dose irradiation affects hematopoietic clonal dynamics and have implications for design of conditioning regimens for transplantation purposes.

Due to high frequency of side effects caused by high-dose total body irradiation (TBI) the nonmyeloablative regimen together with cytotoxic agents is currently used especially for elderly patients. However, immediate and long-term effects of low-dose irradiation used in allogeneic transplantation on stem cells is less well known. We have studied the effect of low-dose 3 Gy TBI on the number of hematopoietic stem cell (HSC) clones contributing simultaneously to granulocyte production in rhesus macaque. The number of clones after 3 Gy TBI decreased markedly by 2 to 3 weeks after 3 Gy TBI, followed by a period of clonal instability, and recovery to almost pre–3 Gy TBI clonal diversity. The clones accounting for this recovery contributed before 3 Gy TBI, suggesting the profound initial impact of TBI was on a pool of progenitor cells, whereas most of the more primitive HSCs remained unaffected and were able to again contribute to hematopoiesis after recovery. Clonal fluctuation may indirectly suggest the presence of short-term/long-term HSC populations in rhesus macaque bone marrow as reported in a mouse model. The results indicate that even low-dose irradiation affects hematopoietic clonal dynamics and have implications for design of conditioning regimens for transplantation purposes.

Background:

Chabannon: Sanofi SA: Other: Travel Support, Research Funding, Speakers Bureau; Bellicum Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees, Other: Travel Support, Speakers Bureau; Novartis: Speakers Bureau; BMS/Celgene: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; Terumo BCT: Speakers Bureau; Miltenyi Biotech: Research Funding; Fresenius Kabi: Research Funding; EBMT: Membership on an entity's Board of Directors or advisory committees. Blaise: Jazz Pharmaceuticals: Honoraria.

Stoppa: celgene: Other: travel fees, lecture fees; takeda: Other: travel fees. Carlo-Stella:MSD: Honoraria; BMS: Honoraria; Janssen: Other: Travel, accommodations; Boehringer Ingelheim: Consultancy; Genenta Science sr: Consultancy; Sanofi: Consultancy, Research Funding; ADC Therapeutics: Consultancy, Other: Travel, accommodations, Research Funding; Novartis: Consultancy, Research Funding; Servier: Consultancy, Honoraria, Other: Travel, accommodations; F. Hoffmann-La Roche Ltd: Honoraria, Other: Travel, accommodations, Research Funding; Rhizen Pharmaceuticals: Research Funding; Celgene: Research Funding; Amgen: Honoraria; Takeda: Other: Travel, accommodations; Janssen Oncology: Honoraria; AstraZeneca: Honoraria. Chabannon:EBMT: Other: Working Party Chair, Board member; Fresenius Kabi: Other: research support; Miltenyi Biotech: Other: research support; Terumo BCT: Other: speaker's fees; Celgene: Other: speaker's fees; Novartis: Other: speaker's fees; Gilead: Other: speaker's fees, hospitalities; Sanofi SA: Other: research support, speaker's fees, hospitalities. Santoro:Takeda: Speakers Bureau; BMS: Speakers Bureau; Roche: Speakers Bureau; Abb-Vie: Speakers Bureau; Amgen: Speakers Bureau; Celgene: Speakers Bureau; Servier: Consultancy, Speakers Bureau; Gilead: Consultancy, Speakers Bureau; AstraZeneca: Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Arqule: Consultancy, Speakers Bureau; Lilly: Speakers Bureau; Sandoz: Speakers Bureau; Eisai: Consultancy, Speakers Bureau; Novartis: Speakers Bureau; Bayer: Consultancy, Speakers Bureau; MSD: Speakers Bureau; BMS: Consultancy. Blaise:Sanofi: Honoraria; Jazz Pharmaceuticals: Honoraria; Molmed: Consultancy, Honoraria; Pierre Fabre medicaments: Honoraria.

Chabannon: Sanofi SA: Other: research support, speaker's fees, hospitalities; Gilead: Other: speaker's fees, hospitalities; Novartis: Other: speaker's fees; Celgene: Other: speaker's fees; Terumo BCT: Other: speaker's fees; Miltenyi Biotech: Other: research support; Fresenius Kabi: Other: research support; EBMT: Other: Working Party Chair, Board member. Vey:Janssen: Honoraria; Novartis: Consultancy, Honoraria. Blaise:Jazz Pharmaceuticals: Honoraria; Sanofi: Honoraria; Pierre Fabre medicaments: Honoraria; Molmed: Consultancy, Honoraria.

BACKGROUND

Stoppa: takeda: Other: travel fees; celgene: Other: travel fees, lecture fees. Vey:Novartis: Consultancy, Honoraria; Janssen: Honoraria. Chabannon:Novartis: Other: speaker's fees; Celgene: Other: speaker's fees; Sanofi SA: Other: research support, speaker's fees, hospitalities; Gilead: Other: speaker's fees, hospitalities; EBMT: Other: Working Party Chair, Board member; Fresenius Kabi: Other: research support; Miltenyi Biotech: Other: research support; Terumo BCT: Other: speaker's fees. Blaise:Pierre Fabre medicaments: Honoraria; Sanofi: Honoraria; Molmed: Consultancy, Honoraria; Jazz Pharmaceuticals: Honoraria.

Advanced hematologic malignancies in older patients are unmet medical needs due to increased incidence in the aging population and the lack of effective treatment. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) represents the only potential curative option for most of hematologic malignancies. However, two main limitations limited this development for a long period of time. High rates of fatal non-relapse morbidities have been progressively addressed using reduced toxicity conditioning approaches and pushing up the age limit of patients considered for transplant. The search for a donor has been also a high limitation in older population, due to the lower incidence of matched sibling donors in this population. The development of allo-HSCT from a familial haplo mismatch donor (Haplo donor) after a reduced intensity (RIC) or non-myeloablative conditioning (NMAC) has allowed for the recent development of allo-HSCT in older population.

Introduction:allogeneic transplantation (allo-HSCT) is a curative treatment for patients with advanced lymphoma. Haploidentical (haplo-SCT) transplantation extended the accessibility to allo-HSCT, overcoming the issue of donor availability. However, alternative donor allo-HSCT is still considered at higher risk of non-relapse mortality due to the HLA disparity and thus an anticipated higher incidence of GVHD. In this context, the use of a non myeloablative conditioning (NMAC) regimen combined with post transplantation cyclophosphamide (PT-Cy) based GVHD prophylaxis may reduce procedure related toxicity. The aim was to evaluate the toxicity and efficacy of haplo-SCT using NMAC with PT-Cy in advanced lymphoma patients.

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) is a curative option for patients with AML in first complete remission (CR1), but is limited by the procedure-related toxicity and mortality. Thus, one could be reluctant for proceeding to Allo-HSCT in elderly patients, especially in the intermediate ELN risk group. While indications of Allo-HSCT for AML are consensual for younger patients, the benefit in older patients remains a matter of debate. In this analysis of a 10-year single center transplantation program for AML, we aim to show the feasibility and study the benefit of Allo-HSCT for CR1 AML in patients over 60 years of age.

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) is a curative option for hematological malignancies. While many procedures have strongly improved outcome after Allo-HSCT by reducing procedure-related toxicity and mortality, disease recurrence remains the major concern. Prophylactic donor lymphocyte infusion (pDLI) is more and more used after Allo-HSCT, with the goal to reduce relapse incidence but may be limited by the risk of developing Graft versus Host Disease (GVHD). The risk of haploidentical DLI-associated GVHD might be increased as compared to HLA-matched DLI. While curative DLI has been assessed for the treatment of hematological relapse after Haplo-SCT, outcomes related to prophylactic DLI has not been evaluated in the setting of Haplo-SCT using post-transplant Cyclophosphomide.

T-cell replete Haplo-SCT using PT-Cy as GVHD part of prophylaxis has become a valid option to perform allogeneic transplantation in patients without an available HLA-matched donor. The absence of additional ex vivo procedure has participated to its increasing use in the field of transplantation. Because of HLA disparity, bone marrow (BM) was initially chosen as stem cell source, based on previous observations of lower incidences of GVHD after HLA-matched donor transplantation, when using BM graft as compared with G-CSF mobilized peripheral blood stem cells (PBSC). In contrast, use of PBSC may improve engraftment and disease control favoring larger adoption of the procedure. However, the use of PBSC graft for Haplo-SCT remains controversial. We here report our experience of PBSC Haplo-SCT using PT-Cy for hematological malignancies.

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) is a curative option for patients with AML. However, most of them are diagnosed over 60 years of age, limiting in part the possibility of intensive treatment. In addition, advanced age is frequently associated with less probability to identify an available matched related donor among siblings (MSD), decreasing again the access to curative treatment. Unrelated donor (UD) Allo-HSCT is an alternative but is associated with higher morbidity, especially related to higher incidence of graft-versus-host disease. More recently, haplo-identical related donor (HRD) Allo-HSCT was developed as a valid option in the absence of a matched donor; still, many investigators reluctantly consider this alternative in elderly patients, anticipating higher morbidity. We here report a single institution comparative evaluation of our HRD vs MSD or UD Allo-HSCT transplant program for patients >= 60.

No relevant conflicts of interest to declare.

Background

Background:

Introduction:Allogenic hematopoietic stem cell transplantation (SCT) is often the only curative treatment for a part of patients with malignant or benign hematological disorders. The availability of a HLA matched related or unrelated donor remains a major obstacle which can be resolved by the presence of alternative donors, like umbilical cord blood, partially matched unrelated or haploidentical family donors. T cell repleted SCT using haploidentical donors (H-SCT) has considerably improved over the last years due to better control of Graft Versus Host Disease (GVHD) using post-transplantation cyclophosphamide (PTCY).

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

Background: High dose chemotherapy followed by autologous stem cell transplantation (ASCT) remain standard of care in patients with relapsed non Hodgkin’s lymphoma (NHL) and Hodgkin’s lymphoma (HL). This regimen is also proposed as consolidation therapy in patients with poor prognosis aggressive NHL and mantle cell lymphoma in first complete remission (CR). BEAM (BCNU, etoposide, cytarabine, melphalan) is the standard protocol chemotherapy used as conditioning regimen. A few previous studies with Bendamustine replacing BCNU have been reported with promising results. BendaEAM seems to show less toxicity and might improve results in relapsed Hodgkin/non-Hodgkin’s lymphoma (R-HL/NHL) patients.

Acute myeloid leukemia (AML) is the most common acute leukemia diagnosed in adults, characterized by significant heterogeneity in terms of biology and clinical outcome. Improvements in sequencing technologies have led to the discovery of frequent somatic mutations in epigenetic modifiers, placing epigenetic deregulation in the center of AML. Yet, the global view and the impact of this deregulation on disease characteristics are under investigation.

Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) is the only curative treatment for many patients with hematologic disease. Unfortunately, the transplanted immune system can reacts against the patient, inducing Graft-versus-Host Disease (GvHD). GvHD is the leading cause of treatment related mortality and morbidity following HSCT and is expected to increase in the forthcoming years. Pathophysiology and long-term determinants of GvHD are still largely unknown.

Introduction:Sickle cell disease (SCD) is one of the most common genetic disorders in the world and despite advances in best supportive care it remains a disease with high risk of morbidity and mortality. Allogeneic hematopoietic stem cell transplantation (alloHSCT) is the only curative treatment modality. Reports of adults are limited because of the lack of suitable matched donors and the high treatment related toxicity (TRT) after conventional myeloablative conditioning regimen resulting from the accumulated disease specific end organ damage. Initial reports of non myeloablative conditioning regimen showed disappointing results because of the high risk of graft failure (GF) and disease recurrence. Recently, the development of reduced toxicity conditioning regimen (RTC) with lower TRT and the use of haploidentical family donors has widened the applicability of alloHSCT. Since SCD patients (pt) are highly alloimmunosized, donor HLA-specific antibodies (DSA) are often detectable and might compromise engraftment in HLA mismatched transplants.

Inhibitory killer immunoglobulin-like receptors (KIR) negatively regulate natural killer (NK) cell–mediated killing of HLA class I-expressing tumors. Anti-KIR treatment resulted in long-term survival in SCID mice inoculated with autologous AML cells (Romagne et al. Blood 114: 2667, 2009). Lack of KIR-HLA class I interactions has been associated with a markedly reduced relapse risk and increased survival in patients with acute myeloid leukemia (AML) upon stem cell transplantation from KIR-mismatched haploidentical donors (Ruggeri et al. Blood 2007).IPH2101 is a fully human monoclonal, hybridoma manufactured antibody (mAb) targeting the KIR2D receptors on NK cells. Results of a dose escalation phase 1 trial conducted in elderly patients with AML in complete remission (CR) indicated a satisfactory safety profile. The maximal tolerated dose was not reached at 3mg/kg. There was a clear correlation between drug exposure and KIR occupancy. Survival was consistent with a dose-related anti-leukemia effect (Vey et al. Blood 120: 4317, 2012. An extension of this phase 1 study was conducted in order to further assess the safety and efficacy of repeated monthly cycles of IPH2101 given at doses producing continuous KIR occupancy. The results of this extension are reported here.This was an open-label, multicenter extension trial to the phase 1 dose-escalation study, conducted in 12 patients ≥ 60 years with an AML in CR. They were to receive up to 6 monthly cycles of IPH2101 at doses of 1 or 2 mg/kg administered as an IV infusion over one hour. Age £ 80, ineligibility for allogeneic cell transplantation, recovery from toxicities of prior chemotherapies and a time lapse ≤ 60 days since the last consolidation cycle were required.Twelve patients were enrolled (median age 71 years; range: 64-75). All were in CR after a median of 3 consolidation cycles (range: 1-6). Seven patients received 6 cycles of IPH2101 and the others between 1 and 3 cycles. Premature discontinuation was due to relapse of AML in four patients and to an unrelated adverse event in one patient (worsening of heart failure).Thirty one related adverse events (AEs) occurred in 83% of the patients. No grade 4 related AEs was reported. Four patients had transient grade 3 related AEs: lymphopenia (4) and amylase/lipase increased (1). Related AEs that were observed in more than one patient included lymphopenia (4), pruritus (3) and erythema/ rash (3). There was no suggestion of a dose effect for safety.The first 6 patients received a dose of 1 mg/kg. At this dose level, full KIR2D occupancy (>95%) was not sustained during monthly cycles in 3/6 patients. Six additional patients received 2 mg/kg with continuous full KIR2D occupancy in all patients. After treatment discontinuation, full occupancy persisted for a median of 1 month (range: 1-3).Median follow-up is 62 weeks at the time of this analysis, 5/12 patients were still alive including 1 in continuous CR All deaths were secondary to AML relapse.Analyses are ongoing to document the impact of IPH2101 treatment on various hematological and immunological parameters in serum (cytokine levels) and in peripheral blood (minimal residual disease), distribution of lymphocytes subsets, as well as phenotypic and functional analysis of NK cells.The safety of IPH2101 was confirmed in this extension cohort assessing repeated monthly cycles leading to full and sustained KIR occupancy for more than 6 months.Vey: Innate Pharma: Honoraria; Bristol Myers Squibb: Honoraria. Andre:Innate-Pharma: Employment, Equity Ownership. Calmels:Innate-Pharma: Employment, Equity Ownership. Zerbib:Innate Pharma: Employment. Buffet:Innate-Pharma: Employment.

Inhibitory killer immunoglobulin-like receptors (KIR) negatively regulate natural killer (NK) cell–mediated killing of HLA class I-expressing tumors. Anti-KIR treatment resulted in long-term survival in SCID mice inoculated with autologous AML cells (Romagne et al. Blood 114: 2667, 2009). Lack of KIR-HLA class I interactions has been associated with a markedly reduced relapse risk and increased survival in patients with acute myeloid leukemia (AML) upon stem cell transplantation from KIR-mismatched haploidentical donors (Ruggeri et al. Blood 2007).

To investigate the impact of pre-transplant CMV serostatus of donor or recipient on outcome of patients undergoing allogeneic hematopoietic stem cell transplantation (Allo-SCT) for Multiple Myeloma (MM).We retrospectively followed 99 consecutive patients who underwent reduced-intensity conditioning (RIC) Allo-SCT for MM in our cancer centre at Marseille between January 2000 and January 2012. Based upon CMV serostatus, patients were classified as low risk (donor [D]-/recipient [R]-) 17 patients (17.1%), intermediate risk (D+/R) 14 patients (14.1%), or high risk – either (D-/R+) 31 patients (31.3%) or (D+/R+), 37 patients (37.3%).Cumulative incidence of CMV reactivation was 39% with a median time of 61 days (26–318). Three patients (3%) developed CMV disease. Two factors were associated with CMV reactivation: CMV serostatus group (low: 0% vs intermediate: 29% vs high: 50%; p=0.001) and the presence of grade II–IV acute GvHD (Hazard Ratio: HR=2.1 [1.1–3.9]). Thirty-six of the 39 patients (92%) with CMV reactivation did not present positive detection of CMV after a 21-day median duration preemptive treatment with ganciclovir. Cumulative incidence of day 100 grade II–IV acute GvHD, 1-year chronic GvHD and day 100 transplantation related mortality (TRM) were 37%, 36% and 9%, respectively. CMV reactivation and serostatus were not associated with increased GvHD and TRM or short survival. Only the presence of acute GvHD as a time dependent variable was significantly associated with increased TRM (p=0.005). Two-year overall and progression free survival were 56% and 34%, respectively.Donor and recipient CMV serostatus and acute GvHD are independent factors for increased CMV reactivation in high-risk MM patients undergoing RIC Allo-SCT. However, we did not find any influence of CMV reactivation on post transplantation outcome. CMV monitoring and pre-emptive treatment strategy could in part explain these results. Novel prophylactic measures such as immunotherapy and drug prophylaxis need to be considered in this specific group of patients, warranting further prospective studies.CR, complete remission; VGPR, very good partial remission; PR, partial remission, PD, progressive disease; Flu, fludarabine; Bu, busulfan; ATG, antithymocyte globulin; CR, complete remission; VGPR, very good partial remission; PR, partial remission, PD, progressive disease; MRD, matched related donor; URD, unrelated donor; GVHD, graft versus host disease; CSA, cyclosporine A; MMF, mycophenolate mofetyl.No relevant conflicts of interest to declare.

Cryopreservation of Hematopoietic Stem Cells (HSC) is a standard procedure for both allogeneic cord blood and autologous HSC transplantation. PBSC are routinely used for the latter; the graft is usually thawed bedside and shortly reinfused without any manipulation. Infusion-related toxicity is usually mild, although severe reactions have been reported, mostly in patients with co-morbidities at baseline. Indeed, cell viability decreases over time after thawing. Washing the thawed graft was reported to improve both clinical tolerance and cell viability. It is however not routinely carried out as it is time-consuming and may result in clinically relevant HSC loss. The fully automated solution SmartWash™ (Biosafe, Switzerland) is designed to wash a thawed graft in a clinical setting. We report here its validation through an international, prospective, multicenter trial.A pre-clinical and a clinical study were designed and sequentially carried out in 4 European Centers (Basel, Florence, Marseille and Murcia) to assess the reproducibility and feasibility of the SmartWash™ system. During the pre-clinical phase, 10 PBSC Units/Center (total: 40 procedures) from patients who permanently lost the indication to the transplant were thawed and split in two aliquots: one was washed and the other was left unmanipulated. A shared protocol targeted to assess recovery, viability and stability of thawed PBSC at different time-points was developed; in particular, an ISHAGE-modified flow cytometry method was implemented in order to improve the evaluation of thawed CD34+ cells. Clinical grade hydroxyethyl starch (Voluven® 6%,Fresenius, Germany) was validated and used as washing solution. Following the validation of the method, a prospective, observational clinical trial was started after approval of the competent Authorities. Twenty-five consecutive patients across the 4 Centers underwent an autologous PBSC transplantation according to the local practice and indications; all the grafts were washed by Smartwash™. Cellular recovery, clinical tolerance and transplant outcome were collected according to a shared protocol.All the washing procedures were completed without any laboratory problems in the participating centers. In the pre-clinical study the recovery of total CD34+ cells in the washed product was 101.2%±27.0 as compared to the pre-freezing value, with a viability of 87.2%±10.0, always showing an advantage over the unwashed samples at all considered time-points (0,1,4 and 24 hours after washing, respectively, Fig. 1). The workload of the procedure, as compared to the unwashed process, resulted in a 40 minutes prolongation for each thawed product, among which only 12 minutes were devoted to active time work, whilst 28 minutes consisted in automatic processing.In the clinical trial, patients received a median of 4.32×106 CD34+ cells/Kg (1.32–27.92), as measured before cryopreservation. Recoveries of both total and viable CD34+ cells after thawing and washing are reported in the Fig. 2, replicating the results of the pre-clinical study. Infusion-related toxicity was negligible (1 grade-1 toxicity out of 25 patients). All patients achieved both neutrophil and platelet sustained engraftment at a median of 11 (9–19) and 13 (7–19) days, respectively.SmartWash™ is an efficient and safe automated method to wash thawed PBSC grafts in a closed, clinical-grade system. Its feasibility and reproducibility was confirmed in an international, multicenter, clinical study. The washed product is stable and clinically feasible for several hours after thawing.Saccardi: Biosafe SA: Research Funding. Boieri:Biosafe SA: Research Funding. Loutan:Biosafe SA: Employment.

Abstract 3164

Abstract 4496

Relapse remains the main issue after allogeneic stem cell transplantation (Allo-SCT) in high risk Multiple Myeloma (MM) patients. The aim of this study is to assess the anti-myeloma effect of lenalidomide followed by donor lymphocyte infusion (DLI) as adoptive immunotherapy after transplantation.Twelve patients with refractory and high risk myeloma were analyzed. Median age at transplantation was 56 years (46–64); 6 patients (50%) received lenalidomide before Allo-SCT. All patients received a RIC including Fludarabine 30 mg/m2 5 days, ATG 2,5 mg/kg for 2 days and Busilvex 3.2 mg/kg/day (3 days in 6 patients and 2 days in 6 patients). All but one received peripheral blood stem cells (PBSC). Donor was HLA id in 6 patients and matched unrelated in 6 patients. It is our standard long term practice to consider post-transplant DLI in patients with progressive or persistent disease after day 100 if no GVHD signs were evident. In 2010, we introduced the use of lenalidomide after day 100 in patients with MM presenting the same characteristics. Doses ranged from 10 to 25 mg/day. Lenalodomide treatment could be completed with DLI, administered afterward, at least after 2 cycles.The median time between Allo-SCT and lenalidomide was 10 months (3–38). The median initial dose of lenalidomide was 15 mg (10–25). Patients received a median of 6 cycles (1–10). Nine patients (60%) received an escalating dose of DLI; 1 × 107/Kg of CD3+cells for the first DLI and 1 × 108/Kg of CD3+cells for the second DLI. One patient with GVHD (after tapering of the cyclosporine A and only after 10 days of lenalidomide) and two patients with progressive disease after lenalidomide did not receive DLI.The toxicity related to lenalidomide was mainly haematological (grade II in 4 patients (33%) and grade I in 3 patients (25%); 7 patients (58%) had moderate asthenia. One patient developed a reversible renal insufficiency after 10 cycles of lenalidomide, none of our patients developed thrombo-embolism under treatment.At the last follow up, 9 patients are alive and all of them are under ongoing treatment. Four patients achieved complete remission (CR) and five patients partial remission at last evaluation. The 1 and 2 years probability of the progression-free survival (PFS) was 75% and 50% and overall survival (OS) was 83 % and 69% respectively. The median OS was not reached and the median PFS was 23 months.These data show that lenalidomide has an acceptable toxicity. Combination with DLI should be further evaluated in a larger cohort of patients.No relevant conflicts of interest to declare.

The purpose of this study was to assess the results of allogeneic stem cell transplantation (Allo-SCT) after reduced-intensity conditioning (RIC) from an unrelated donor in patients with high-risk multiple myeloma (MM) in a single centre. From January 2007 to January 2011 we consecutively transplanted 40 patients with MM.Seventeen (43%) (Group 1) and 23 patients (57%) (Group 2) had unrelated and related donor respectively. The median age was 48 years (39–63) in the first group and 56 years (40–67) in the second group (P=0.0769).Thirty nine patients (98%) received one or more autologous transplantation. Ten patients (Group 1: N=5 (29%); Group 2: N=5 (22%)) were transplanted within whereas 30 patients were treated beyond (Group 1: N=12 (71%); Group 2: N=18 (78%) the first line treatment strategy. The disease status at transplantation was Complete Remission (CR) or VGPR in (35%) vs (43%), Partial remission (PR) in (59%) vs (52%) and Progression or Refractory Disease in (6%) vs (4%) (PD/RD) in the first and second group respectively (p=0.1770). Graft was peripheral blood stem cells (PBSC) in all patients in the related donor group and in 14 patients (82%) in the second group, the other 3 patients (18%) receiving marrow.Thirty-three patients (Group 1: N=14 (82%); Group 2: N=19 (83%) were treated with a RIC based on Fludarabine (30mg/m2/d × 5); Busulfan (4 mg/kg/d p.o. or 3.2 mg/kg/d IV over 2 to 3 days) and rabbit ATG (2.5 mg/kg/d × 2). Seven patients received Fludarabine (25mg/kg/d for 3 days) and 2 Gys total body irradiation (TBI).Post-graft immunosuppression consisted of cyclosporine (CSA) alone in 26 patients (65%), CSA and mycophenolate mofetil in 14 patients (35%).The median follow-up was 22 months (1–49). None of our patient experienced a graft rejection. The cumulative incidence of grade II-III acute graft versus-host disease (GVHD) tended to be higher after unrelated donor graft (41% vs 17%) (p= 0.12). The cumulative incidence of chronic GVHD was no different between the first and second group (24% vs 30% respectively). At last follow up 24 patients (group 1: N=11 (65%); Group 2:N=14 (61%) were still alive of whom 17 are in CR (group 1: N=9 (53%); Group 2:N=8 (35%), 5 in PR (group 1: N=1 (6%); Group 2:N=4 (17%)and 2 in progressive disease (group 1: N=1; Group 2:N=1).The estimated probability of non relapse mortality (NRM) at day 100 was 0% in the two groups and not statically different at 2 years. (12% vs. 22% (P=0.4)) Also 2 year overall and progression-free survivals were not statiscally different after unrelated and related donor transplants (59% vs. 66% (P=0.110) and 42% vs. 44% (p=0.241)). The incidence of acute GVHD, OS, PFS and NRM were not significantly different between the two groups.Our experience, although limited, supports that high risk MM patients can benefit from a RIC Allo-SCT with unrelated donor when a HLA matched sibling donor is not available conducting to acceptable and comparable outcomes. This deserves further analysis in larger populations.No relevant conflicts of interest to declare.

No relevant conflicts of interest to declare.

Abstract 4545

Abstract 4513