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We conducted a prospective evaluation of cord blood (CB)–derived adoptive cell therapy, after salvage chemotherapy, for patients with advanced myeloid malignancies and poor prognosis. Previously, we reported safety, feasibility, and preliminary efficacy of this approach. We present updated results in 31 patients who received intensive chemotherapy followed by CB infusion and identify predictors of response. To enhance the antileukemic effect, we selected CB units (CBU) with shared inherited paternal antigens and/or noninherited maternal antigens with the recipients. Twenty-eight patients with acute myeloid leukemia (AML), 2 with myelodysplastic syndrome, and 1 in chronic myeloid leukemia myeloid blast crisis were enrolled; 9 had relapsed after allogeneic transplant. Response was defined as

Adoptive immunotherapy has shown efficacy in patients with relapsed/refractory acute myelogenous leukemia (AML). We conducted a prospective evaluation of cord blood (CB)-based adoptive cell therapy following salvage chemotherapy in patients with AML or myelodysplastic syndrome (MDS) and describe the safety and early outcomes of this approach. To enhance the antileukemic effect, we selected CB units (CBUs) with a shared inherited paternal antigen (IPA) and/or noninherited maternal antigen (NIMA) match with the recipients. Furthermore, the CBUs had total nucleated cell (TNC) dose

Adolescents and young adults (AYAs) with cancer are a unique group of patients in terms of disease incidence and biology, outcome, and psychosocial needs. This study aims to correlate the risk of graft-versus-host disease (GvHD) and age in a population of children and young adults with acute leukemia undergoing hematopoietic stem cell transplantation (HSCT) in first complete remission (CR).We analyzed the outcome of 153 consecutive children (15 years), AYAs (15-24 years), and adults (25-35 years) with lymphoblastic or myeloid acute leukemia in first CR who underwent HSCT with matched donors after myeloablative conditioning. GvHD prophylaxis was methotrexate and cyclosporine A (CsA) in all patients.The cumulative incidence of grade II-IV acute GvHD (aGvHD) was significantly higher in AYA patients than in children (subdistribution hazard ratio (SHR), 2.04, p = 0.005) or adults (SHR 1.59, p = 0.048). Both gut and skin aGvHD occurred more frequently in AYA patients. Increasing CsA blood levels with age could not fully account for this difference. No difference in terms of grade III-IV aGvHD was observed. Chronic GvHD was more frequent in AYAs (SHR 2.81, p = 0.007) and adults (SHR 2.31, p = 0.033) than in children. No difference in terms of nonrelated mortality and overall survival was observed among the age subgroups.Since GvHD occurrence is strongly correlated to quality of life, specific attention should be paid to AYAs undergoing HSCT. Further studies should investigate the reasons for the excess of GvHD observed in this population.

In absence of available matched-related or unrelated donor (MUD), mismatched unrelated donors (MMUD) and unrelated cord blood (UCB) are both considered to be suitable donors, with similar post-transplant overall survival. In most of these retrospective comparisons, HLA typing of adult donors was performed at eight loci. The aim of this study was to compare the outcome of patients transplanted from UCB (N = 64) with those transplanted from 9/10-HLA MMUD (N = 84) or 10/10-HLA MUD (N = 196). In multivariate analysis, UCB was associated with less Grade II–IV acute GVHD in comparison with MUD (aHR 1.97, 95% CI 1.19–3.27, P = 0.009) and MMUD transplants (aHR 1.79, 95% CI 1.02–3.15, P = 0.042), while the cumulative incidence of chronic GVHD was not significantly different between the three groups. Overall survival (OS), non-relapse mortality, and relapse were not different between MMUD and UCB transplantation, whereas OS was impaired after UCB in comparison with MUD (aHR 0.65, 95% CI 0.43–0.99, P = 0.043). Factors also impacting OS were the donor/recipient CMV serostatus (Donor−/Recipient+ aHR 1.76, 95% CI 1.23–2.52, P = 0.002 compared with D−/R−), the donor/recipient gender combination (Female/Male versus other combinations aHR 1.57, 95% CI 1.11–2.22, P = 0.012) and disease risk (aHR 1.58, 95% CI 1.05–2.38, P = 0.027 for high vs. low risk disease). Our data confirm that UCB and 9/10-HLA MMUD are both relevant alternative options when no 10/10-HLA donor is available. Donor/recipient gender combination and CMV serostatus had a significant impact on survival and may be taken into account, along with donor type, in the setting of MMUD and UCB transplants. Am. J. Hematol. 90:897–903, 2015. © 2015 Wiley Periodicals, Inc.

Natural killer cells are the first lymphocyte subset to reconstitute, and play a major role in early immunity after allogeneic hematopoietic stem cell transplantation. Cells expressing the activating receptor NKG2C seem crucial in the resolution of cytomegalovirus episodes, even in the absence of T cells. We prospectively investigated natural killer-cell reconstitution in a cohort of 439 adult recipients who underwent non-T-cell-depleted allogeneic hematopoietic stem cell transplantation between 2005 and 2012. Freshly collected blood samples were analyzed 3, 6, 12 and 24 months after transplantation. Data were studied with respect to conditioning regimen, source of stem cells, underlying disease, occurrence of graft-versus-host disease, and profiles of cytomegalovirus reactivation. In multivariate analysis we found that the absolute numbers of CD56(bright) natural killer cells at month 3 were significantly higher after myeloablative conditioning than after reduced intensity conditioning. Acute graft-versus-host disease impaired reconstitution of total and CD56(dim) natural killer cells at month 3. In contrast, high natural killer cell count at month 3 was associated with a lower incidence of chronic graft-versus-host disease, independently of a previous episode of acute graft-versus-host disease and stem cell source. NKG2C(+)CD56(dim) and total natural killer cell counts at month 3 were lower in patients with reactivation of cytomegalovirus between month 0 and month 3, but expanded greatly afterwards. These cells were also less numerous in patients who experienced later cytomegalovirus reactivation between month 3 and month 6. Our results advocate a direct role of NKG2C-expressing natural killer cells in the early control of cytomegalovirus reactivation after allogeneic hematopoietic stem cell transplantation.

Pregnancy is the only natural source of anti-HLA immunization. The exact frequency of this immunization remains undetermined as prior studies either used methods with a low sensitivity or were performed late after delivery. We present here the first study on women at delivery evaluating anti-HLA immunization by Luminex. We also attempted to isolate factors influencing immunization, such as soluble HLA-G (sHLA-G) levels and genetic polymorphisms. With Luminex, anti-HLA immunization was observed in 54.4% of the women. As expected, immunization frequency increased with the number of children, reaching 74% in women with >2 deliveries. Among immunized women, strong cytotoxic Ab (as detected by Complement Dependent Cytotoxicity) were associated with a lower level of sHLA-G. None of the studied polymorphisms influenced immunization rate in the whole cohort. Among 94 first pregnant women with no history of miscarriage, the -174 IL-6 gene promoter mutation (G/C) appeared more frequently in non immunized women (69% vs. 45% in immunized ones, p = 0.02). Lastly, the occurrence of a miscarriage before the first live delivery significantly decreased immunization. These results may help to understand mechanisms of pregnancy induced immunization. They also have an impact in the management of previous pregnant women requiring organ or hematopoietic stem cell transplantation.

International audience; Graft failure is a major complication after unrelated cord blood transplantation. Presence of HLA-antibodies before cord blood transplantation may impact graft failure. To analyze the effect of anti-HLA antibodies on unrelated cord blood transplantation outcomes, we analyzed 294 unrelated cord blood transplant recipients after reduced intensity conditioning regimen. The majority of the patients (82%) were transplanted for malignancies, 60% with double-unrelated cord blood transplant, 63% were HLA mismatched. Retrospectively, pre-unrelated cord blood transplant serum was tested for HLA-Ab using Luminex™ platform. Results were interpreted as mean fluorescence intensity (MFI) against donor-specific mismatch. Among 62 recipients (23%) who had anti-HLA antibodies before unrelated cord blood transplant, 14 patients had donor specific anti-HLA antibodies (DSA) (7 were donor-specific anti-HLA antibodies for single unrelated cord blood transplant and 7 for double unrelated cord blood transplant). Donor specific anti-HLA antibodies threshold ranged from 1620-17629 of mean fluorescence intensity (MFI). Cumulative incidence of Day-60 neutrophil engraftment was 76%: 44% for recipients with donor specific anti-HLA antibodies and 81% in those without donor specific anti-HLA antibodies (P=0.006). The cumulative incidence of 1-year transplant related mortality was 46% in patients with donor specific anti-HLA antibodies and 32% in those without antibodies (P=0.06). The presence of donor specific anti-HLA antibodies was associated with a trend for decreased survival rate (42% vs. 29%; P=0.07). Donor specific anti-HLA antibody in recipients of unrelated cord blood transplant is associated with graft failure and decreased survival. Patient's screening for donor specific anti-HLA antibodies before unrelated cord blood transplantation is recommended before choosing an HLA mismatched cord blood unit. Whenever possible it is important to avoid selecting a unit for which the patient has donor specific anti-HLA antibodies.

Background: CBU selection for transplant is based on HLA match and potency/quality determinations associated with the speed of hematopoietic engraftment. Total Nucleated Cell (TNC) count, viable (v) CD45+ and CD34+ cells and Colony Forming Units (CFU) have been used as surrogate markers of hematopoietic stem cells. However, besides TNC measurements, most of the assays still have poor standardization among laboratories. The NCBP has manufactured over 60,000 clinical CBU (as of 07/2018). Of those, 32,413 CBU had pre-cryopreservation samples tested prospectively (period: 03/2008-04/2018) using validated assays for TNC (Sysmex XE-2100), vCD45+ and vCD34+ cells (single platform flow cytometry using ISHAGE strategy and 7-AAD for viability) and HRDI-CFU assay (High Resolution Digital Imaging CFU strategy, Albano et al, Blood 2011;118:485). Among those, 754 CBU have been shipped to US Transplant Centers (TC) for single or double unit transplants. Prior to release for clinical use, CBU had also quality/potency tests conducted prospectively in post-thaw samples from attached segments, using the same assays. Aims and study design: 1) to evaluate the quality/potency of 754 NCBP CBU provided for transplantation (period: 03/24/2008-12/31/2017) by pre-cryopreservation characteristics, post-thaw segment evaluations and their correlations, and 2) to assess their impact on engraftment in two patient subsets after myeloablation: cohort 1, N=99 patients (pts), who received single CBU transplants, and cohort 2, N=139 pts with double unit grafts, who engrafted with the NCBP CBU. All laboratory evaluations were performed by NCBP. Clinical data were provided by CIBMTR; only pts with information on time to engraftment (ANC>500) and established donor chimerism were analyzed. Results: The 754 CBU selected by TC had quality/potency parameters showing a strong correlation between cell numbers and function pre-cryopreservation and post-thaw (Table 1; Pearson correlation). Segment post-thaw median recoveries were 73% (SD: 21.8%) for vCD45+ cells; 69% (SD: 20.7%) for vCD34+ cells and 60% (SD: 22.9%) for CFU, with a strong correlation between pre-cryopreservation and post-thaw values (Pair T Test: 0.74; 0.87 and 0.79, respectively; all P500 was 16 days (range: 7-45). Cohort 2 had 85% adult pts with average age 33 years, average weight 69 kg (range: 11-129) and median time to ANC>500 19 days (range: 6-68). Engrafting CBU median pre-cryopreservation TNC was 2.6x10^7/kg and vCD34+ 1.4x10^5/kg. CBU pre-cryopreservation and segment quality/potency markers showed very good correlation also (Table 1). In cohort 1 all pre-cryopreservation cell doses and segment vCD34+ and CFU correlated with neutrophil engraftment (Table 2). Pts with earlier engraftment (≤16 vs >17 days) received CBU with significantly higher doses of TNC, vCD34+, vCD45+ and CFU (pre-cryopreservation and post-thaw tests). In the recipients of double CB transplants, the engrafting CBU TNC and vCD45+ cell doses did not correlate with neutrophil engraftment, but the vCD34+ and CFU did (Table 2). Similarly, pts with earlier engraftment (≤19 vs >20 days) received CBU with higher doses of vCD34+ and CFU, but not TNC or CD45+. Conclusions: Prospective testing of HPC, CB products manufactured by a single CB Bank over a 10-year period and provided for transplant show consistent, highly significant correlation among pre-cryopreservation graft characteristics and segment measurements. Pre-cryopreservation values were predictive of time to ANC>500 in myeloablated recipients; in those cases, segment evaluation did not improve the correlations. Notably, vCD34+ and CFU correlations are consistently maintained after cryopreservation and thaw ensuring a HPC, CB product with reliable performance for transplantation, cell therapies or use with expansion technologies. Disclosures No relevant conflicts of interest to declare.

International audience; BACKGROUND: Cytomegalovirus (CMV) has a role in chronic rejection and graft loss in kidney transplant (KTx) and lung transplant (LTx) recipients. In addition, donor CMV seropositivity is an independent risk factor for renal graft loss. The anti-CMV response might modulate this risk. Expression of programmed cell death 1 (PD-1), a receptor involved in viral-specific T-cell exhaustion, is influenced by a single nucleotide polymorphism called PD-1.3 (wild-type allele G, variant allele A). METHODS: We performed a retrospective study to assess the impact of PD-1.3 on graft outcome in donor CMV seropositive (D+) and donor CMV seronegative (D-) KTx and LTx. We also performed a case-control study to evaluate the anti-CMVpp65 response according to genotype. RESULTS: PD-1.3 was determined in 1,119 KTx and 181 LTx. In 481 D+ KTx, A allele carriers (24%) experienced significantly less graft failure compared with GG carriers (p = 0.001). Multivariate analysis showed that this association was independent of donor and recipient age, acute rejection episodes, and number of human leukocyte antigen mismatches (hazard ratio, 0.381; 95% confidence interval, 0.209-0.696; p = 0.002). Analysis in 85 D+ LTx showed similar results: A allele carriers had better survival (hazard ratio, 0.302; 95% confidence interval, 0.128-0.716; p = 0.006) and greater 6-month forced expiratory volume (71% ± 17% vs 54% ± 16%, p = 0.001). In D- recipients, PD-1.3 did not affect KTx or LTx outcome. Finally, AA recipients had a stronger anti-CMVpp65 T-cell response than matched GG recipients (p = 0.003). CONCLUSIONS: The A variant allele in PD-1.3 single nucleotide polymorphism improved graft survival in kidney and lung transplant recipients receiving grafts from CMV-positive donors.

Human leukocyte antigen (HLA) and non-HLA immunogenetic factors impact the outcome of hematopoietic stem cell transplantation (HSCT). While in early studies of cord blood transplantation (CBT) the major histocompatibility complex immunogenetic barrier was considered less important in some cases than in HSCT from unrelated donors, and preference was given to the number of cells infused as a success criteria, the most recent data emphasize the critical role for HLA matching. This affects the choice of strategy for immunogenetic typing for cord blood banking. Other immunogenetic systems may be of interest in the context of CBT and warrant further investigation. Finally the role of antibodies is also discussed as well as the overall immunogenetic assessment (HLA and beyond HLA). It will provide a tailored and personalized choice of the best donor for each individual patient.

Mobilized peripheral blood has become the predominant stem cell source for allogeneic hematopoietic cell transplantation. In this retrospective single center study of 442 patients with hematologic malignancies, we analyzed prognostic factors for long-term survival after peripheral blood stem cell transplantation from HLA-matched related or unrelated donors. To account for disease/status heterogeneity, patients were risk-stratified according to the Disease Risk Index. Five-year overall survival was similar after transplants with matched related and unrelated donors (45% and 46%, respectively; P=0.49). Because donor age ≥60 years impacted outcome during model building, we further considered 3 groups of donors: matched unrelated (aged

International audience; Hyperacute rejection is an early complication of transplantation, caused by the presence in the recipient of pre-formed donor-directed antibodies (Ab). We report a case of fatal early graft dysfunction after lung transplantation in a female recipient with no anti-HLA Ab detected before transplantation. Further immunologic studies revealed the presence, in the pre-transplant serum, of low-titer anti-HLA Ab directed against the donor's HLA, detectable only by flow-cytometry screening for panel-reactive antibodies. This observation emphasizes the importance of sensitive Ab detection in patients awaiting lung transplantation. Women should be specifically targeted for such sensitive Ab detection.

Abstract 2000 Background: Over the past 20 years, allogeneic hematopoietic cell transplantation (HCT) has been increasingly performed with peripheral blood stem cells (PB) and gained benefit from better HLA-typing. Similar long-term survival has been suggested after HLA-matched related and unrelated donor HCT. Till now, the optimal strategy for donor selection is still controversial. We evaluated the impact of donor type (10/10 HLA-matched unrelated (MUD) vs. matched related (MRD)) and other donor traits on long term outcomes of patients with hematologic malignancies after PB HCT. Patients and Methods: We analyzed outcomes of 442 consecutive patients with hematologic malignancies who were transplanted with PB either from MUD (n= 164) or MRD (n=278) at our center from 01/2000 to 12/2010. Median patient age was 48 years (range 7–68). Diseases included 122 acute myelogenous leukemias, 62 non-Hodgkin lymphomas, 60 myelodysplastic syndromes, 57 multiple myelomas, 40 acute lymphoblastic leukemias, 37 myeloproliferative disorders, 29 Hodgkin diseases, 20 chronic myeloid leukemias and 15 chronic lymphocytic leukemias. Two-third of patients underwent HCT following reduced intensity conditioning. Graft-versus-host disease (GVHD) prophylaxis consisted mostly in cyclosporine plus MMF or methotrexate. ATG was used in 19% of HCT. We assessed the impact of donor factors (type, age, gender, CMV serologic status and ABO group) on chronic GVHD (cGVHD), relapse, non relapse mortality (NRM) and overall survival (OS). Concerning donor age, as the upper age limit for voluntary PB donation was usually 60 years, we completed our analysis by performing 3 groups according to donor type and age (MUD, MRD Donors: Median donor age was 40 years (range 18–72). Most young donors were MUD ( Results: The median follow-up (FU) was 36 months (range 2–133) and 25% of patients had a FU of at least 60 months. The cumulative incidence (CIf) of cGVHD at 2 years was 58%. In multivariate analysis, sex mismatch (female > male) increased risk of cGVHD (HR: 1.41 [95% CI 1.05–1.88], P=.02) while MRD≥60y resulted in lower risk (HR: 0.48 [0.25–0.94], P=.031). Donor type by itself did not impact on cGVHD (58% with MRD and 59% with MUD). At 5 years, the CIf of relapse was 34% and was higher with MRD than MUD (39% vs. 24%, P=.038). Adjusted for disease risk, conditioning and infused cells count, only MRD≥60y resulted in significant higher risk of relapse than MUD (HR 2.41[1.26–4.62], P=.008) while MRD Conclusion: Donor/recipient gender parity, donor type and age appeared as significant predictive factors of long term outcomes after HLA-matched PB HCT. Nor donor CMV status nor ABO group seemed to impact on outcomes in our cohort. The selection of a sex mismatched donor (female>male) was associated with significant higher risk of cGVHD. Using PB as graft source, HLA 10/10 MUD provided higher NRM but better disease control and similar OS than MRD. Having combined donor type and age, we observed notable poor outcome (high relapse rate and low OS) for patients transplanted with MRD≥60y in our cohort. Given those results, one may question HCT with old MRD when a younger MUD is available. Disclosures: No relevant conflicts of interest to declare.

Abstract 3135 Background: When an HLA-matched donor is not available for allogeneic Hematopoietic Stem Cell Transplantation (HSCT), the use of an alternative HLA-mismatched stem cell source may be considered. In Europe, HLA compatibility for HSCT is calculated for 10 alleles (HLA-A, -B, -C, -DRB1, -DQB1). The aim of this study was to retrospectively compare outcomes after transplantation from 10/10 HLA-MUD, 9/10 HLA-MMUD and UCB performed at the Hospital Saint Louis (Paris, Fr). Methods: Patients receiving a first allogeneic transplantation from a 10/10, 9/10 UD or UCB from 2000 to 2011 were included. High resolution HLA typing was performed by PCR SSO and SSP for HLA loci: A, B, C, DRB1 and DQB1. The following variables were studied as risk factors for transplant outcomes: disease, disease risk (standard/high risk), age at transplant, gender, donor/recipient sex-matching, ABO matching, donor/recipient CMV status, conditioning regimen, and use of anti-thymoglobulin (ATG). Results: 355 consecutive patients with hematologic malignancies were analyzed. One hundred and ninety-six were transplanted with MUD, 84 with MMUD (mismatches for HLA-A: 16%, -B: 16%, -C: 39%, -DRB1: 8%, -DQB1: 21%) and 75 with UCB (52% with single and 48% with two UCB unit; 87% of all UCB transplants were 4–6/6 HLA-matched). Median patient age was 31 (range: 5–55). Patient characteristics differed between the 3 groups: (i) median age at transplant: 36 in MUD, 31 in MMUD, 22 in UCB (p Cumulative incidences of grade II-IV and grade III-IV acute GvHD disease (aGvHD) were 61% (66% for MUD, 60% for MMUD and 48% for UCB) and 17% (17%, 24% and 15%), respectively. Three-year cumulative incidence of chronic GvHD (cGvHD) was 46% (51% for MUD, 49% for MMUD and 29% for UCB). Three-year NRM was 34% (28% for MUD, 31% for MMUD and 51% for UCB). Graft failure occurred in 15% of UCB patient, 8% in MMUD group and 3% in MUD group (significant difference between UCB and MUD: OR=5.44, 95%CI 1.93–15.3, p=0.001). Multivariate analysis is summarized in table 1. It showed that MMUD tented to have a higher incidence of aGvHD III-IV than MUD and UCB. UCB had a lower incidence of cGvHD than MMUD. No significant effect of HSCT source on NRM was demonstrated. Conclusion: Compared to MMUD, UCB-HSCT induces less cGvHD and non significantly increased NRM. Compared to MUD, MMUD and UCB-HSCT did not result in a clear increase of NRM. UCB and 9/10 HLA-MMUD are both suitable stem cell sources for patients who cannot benefit from 10/10 HLA-MUD transplant. Disclosures: No relevant conflicts of interest to declare.

We have recently showed in a murine bone marrow transplantation (BMT) model that apoptotic cell infusion simultaneously with a bone marrow (BM) graft favors hematopoietic engraftment, and induces a long term tolerance restricted to BM cell allo-antigens. This effect is observed whatever the origin of apoptotic cells and does not lead to the development of autoimmunity or GvHD. This favorable effect on engraftment was obtained without immunosuppressive (IS) drugs. In human BMT, IS treatment is always required. The aim of this study was to investigate the interactions between apoptotic cells and IS drugs, especially since some of them might interfere with other strategies of tolerance induction. The roles of ciclosporin A (CSA), mycophenolate mofetil (MMF), sirolimus (Rapa) and LF150195 (LF), a deoxyspergualin analog, were tested. Recipient Balb/c mice (H2d) were sublethally irradiated (6 Gy) 1 day before receiving 106 BM cells with or without apoptotic cells (5x106 γ-irradiated splenocytes) from donor FvB mice (H2q). Drugs were injected IP: CSA 50mg/kg/d from day 0 (d0) to d14, MMF 20mg/kg/d from d0 to d14, Rapa 1.5mg/kg/d from d0 to d14, and LF 0.625mg/kg/d from d0 to d10. Engraftment was assessed by flow cytometry analysis in peripheral blood. Skin grafts from BM donor mice were performed >70 days after BMT, to assess the influence of IS drugs on our tolerance induction protocol. CSA treatment didn’t favor engraftment in mice receiving BM alone (22% engraftment vs 20% without IS drug, p=NS), and induced only transient chimerism (