ALBERT

Description: Abstract 1811 Poster Board I-837 The prognostic significance of achieving complete remission (CR) in Multiple Myeloma (MM) has finally been accepted. However, available studies have been based on series with a median follow-up around 5 years. This time period is insufficient according to the current life expectation of MM. Aim To establish the real effect of prognosis of the different response categories in a cohort of MM patients treated with autologous stem cell transplantation (ASCT) after long term follow up. Patients and methods Follow-up from diagnosis of 344 MM patients transplanted between 1989 and 1998 has been updated. These patients were previously included in a study aimed at establishing the post-ASCT response significance in MM and to validate the EBMT classification (Br J Haemat 2000;109:438-46). It was possible to update the follow up of 322 patients as at April 2009. At this date 99 patients were alive with a median follow-up form diagnosis of 12.5 years. Response categories and evaluated cases were: i) Complete Response (IF-) (CR), n= 84 ii) near Complete Response (EF-/IF+) (nCR), n= 66 iii) Very good partial response (VGPR) (

Description: The optimal post remission therapy for HR-ALL patients (pts) is not well established. This multicenter randomized trial was addressed to compare three options of post remission therapy in adults with HR-ALL: intensive consolidation plus maintenance chemotherapy (CHT), ALLO or AUTO SCT. Inclusion criteria: one or more of the following: age 30–50 yr., WBC〉25x109/L, t(9;22) or BCR/ABL, 11q23 or MLL and t(1;19). Treatment schedule: 5-drug (VCR, DNR, PDN, ASP, CPM) induction therapy over 5-weeks, followed by 3 1-wk cycles of early intensification CHT including HD-MTX, HD-ARAC and HD-ASP over 3 mo. Patients with an HLA-identical sibling were assigned to ALLO SCT and the remaining were randomized to AUTO SCT or to delayed intensification CHT (the same three cycles given in the post-remission period) followed by standard maintenance CHT (MP+MTX) up to 2-yr. in continuous complete remission (CR). Study period: 1993–2004, 254 pts included, 222 evaluable, 35 hospitals, 131 males, mean (SD) age 29(10) yr, WBC count 60(98) x109/L, early pre-B: 43 (19%), common+pre-B: 113 (51%), T: 66 (30%). Cytogenetics: 161 (73%) valid cases after central review, normal: 67 t(9;22): 37, 11q23: 6 cases, t(1;19): 2, other rearrangements: 49 cases. Response to therapy: CR 183 (82%). Slow response to therapy (〉10% blasts in BM aspirate at day 14 of induction therapy) in 40% of cases. 84 pts were assigned to ALLO SCT, 50 randomized to AUTO SCT and 48 to CHT, 1 pt. removed from protocol after the end of induction. With a median follow-up of 70 mo.(range 24–133), medians for DFS and OS for the whole series were 17 and 23 mo., and 5-yr DFS and OS probabilities were 35±5% and 34±6% (37±6% and 35±6% after removing Ph+ ALL pts from analysis). Groups of ALLO, AUTO and CHT were comparable for the main clinicobiologic characteristics and the rate of response to therapy. Intention-to-treat analysis showed no differences in DFS and in OS for donor vs. no donor comparison as well as for comparison of AUTO vs. CHT. This lack of differences persisted after removing Ph+ ALL pts from the analysis. The same results were observed when the analysis was performed on the basis of effectively treated pts (ALLO SCT performed in 70%, AUTO SCT in 62% and CHT in 73% of the pts). By multivariate analyses, age〉30 yr. was associated with death in induction, slow response to induction therapy and Ph+ ALL were associated with a lower CR, and these three variables had a negative influence on DFS and on OS probabilities. No differences in outcome were found in pts assigned to ALLO SCT or randomized to AUTO SCT or CHT as post-remission therapy in HR-ALL. DFS from ALLO or AUTO SCT or from intensification therapy was also identical in effectively treated patients. Adults with Ph+ ALL or slow response to induction therapy have been identified as a very-high-risk ALL subgroup for whom specific or experimental therapies are justified.

Description: Introduction: Diffuse large B-cell lymphoma (DLBCL) is one of the most common malignant neoplasms in elderly patients, potentially curable when optimum treatment is administered. The combination of rituximab with CHOP chemotherapy (R-CHOP) is considered standard for these patients, but randomized studies published to date are limited to the range of age from 60 to 80 years, so that in patients over this age treatment election is not so clear, usually opting for palliative treatment or a "full" treatment at a reduced dose. This retrospective study is primarily aimed to analyze the influence of the type of treatment and comorbidity scales in overall survival (OS) of a large series of patients 〉80 years with aggressive B-cell lymphoma. Methods: Eligible patients were aged ≥ 80 years, diagnosed of DLBCL, follicular lymphoma grade 3B or transformed lymphoma. The main patient characteristics were obtained retrospectively from the medical records, including a complete geriatric assessment (CGA, "comprehensive geriatric assessment") and the Charlson comorbidity index. The Ethics Committee of the University Hospital of Salamanca approved the study. Results: 288 patients from 19 GELTAMO hospitals were registered in the study, of which 234 (60% women) were evaluable and have been included in this preliminary analysis. The median age was 84 years (80-94) and the vast majority (94%) were DLBCL. According to the Charlson index, 65% of patients were low-intermediate risk, and according to CGA, 63% of patients were considered "fit". A higher proportion (60% v 44%, p = 0.03) of patients with low or intermediate comorbidity index were treated with a curative intent (CHOP +/- rituximab), as compared with patients with high or very high index. With a median follow up of 41 (range 9-142) months, the median OS was 11.5 months (33% estimated at 3 years). The median OS for patients treated with R-CHOP-like (N=96) was 35.3 months, significantly better (p

Description: Introduction: Relapses after front-line therapy for Burkitt lymphoma/leukemia (BL) are unfrequent, and there is scarce information about the best rescue strategy for these patients. The objective of this study was to evaluate the incidence of relapse, salvage treatment and prognosis after relapse in patients with BL treated with two consecutive Spanish protocols. Patients and methods: Retrospective study of patients diagnosed with BL in 40 Spanish hospitals betwen January 1997 and October 2014 treated with first line chemotherapy according to protocols PETHEMA LAL-3/97 (specific chemotherapy without rituximab) and BURKIMAB (rituximab plus specific chemotherapy). The demographic, clinical and biological characteristics were collected at the time of diagnosis and at relapse, as well as the salvage treatment and outcomes. Results: 233 patients were diagnosed with Burkitt lymphoma (n=150) or leukemia (n=83) and received first-line therapy according to PETHEMA LAL-3/97 (n=53) and BURKIMAB (n=180) protocols. Baseline characteristics at diagnosis are described in Table 1. A total of 26 patients relapsed, 11 (28%) treated with PETHEMA LAL-3/97 protocol and 15 (10%) with BURKIMAB protocol (p=0.009). The cumulative incidence of relapse at 10 years was 27% (95% CI, 12%-42%) in PETHEMA LAL-3/97 protocol vs.16% (95% CI, 4%-28%) in BURKIMAB protocol (p= 0.013) (Figure 1). Time to relapse was shorter in PETHEMA LAL-3/97 protocol (median of 3.7 months) vs. BURKIMAB protocol (6.3 months), but it was not significant (p=0.506). No differences were observed in relapse incidence between Burkitt leukemia and Burkitt lymphoma in PETHEMA LAL-3/97 protocol (6/31 vs. 5/22, p=1) and BURKIMAB protocol (7/41 vs. 8/107, p=0.124). Out of 15 patients in whom rescue treatment strategy was evaluable, 12 received chemotherapy with high-dose methotrexate and/or cytarabine (4 of the them followed response, CR in 2, followed by SCT in the 2 patients achieving PR [autologous in one and allogeneic SCT in the other]), and the remaining 3 patients received DA-EPOCH-R (n=1, achieving CR), R-ICE (n=1, no response) and paliative care (n=1). At the time of the analysis, only 3 patients are alive. Median overall survival after relapse was 3 months (95% CI, 0.9-5.1) for PETHEMA LAL-3/97 relapsed group and 3.6 months (95% CI, 0.1-7.1) for BURKIMAB relapsed patients group. Conclusions: Patients with Burkitt leukemia/lymphoma treated with specific immunochemotherapy have lower probability of relapse compared with those treated with specific chemotherapy without rituximab. In our series, the most frequent regimens administered for the treatment of relapsed patients were based in high-dose methotrexate and/or cytarabine. The prognosis of relapsed Burkitt leukemia/lymphoma is poor, independently of the type of rescue therapy. Supported by grants RD12/0036/0029 (RTICC, FEDER), Instituto Carlos III, Spain. Disclosures No relevant conflicts of interest to declare.

Description: Myeloablative transplant has been investigated in poor prognosis indolent lymphoma; although recurrence rate is low it is associated with high mortality; the use of non-myeloablative conditioning regimens could reduce TRM maintaining the GVH effect. Up to May 2006, 35 patients with follicular NHL received a Non Myeloablative related allogeneic according to two prospective multicenter trials; conditioning regimen consisted of Fludarabine 150 mg and Melphalan 70–140 mg. GVHD prophylaxis consisted of CSA plus short-course MTX. All patients received filgrastim-stimulated peripheral blood stem cells from a HLA related identical donor. Median age at transplant was 50 years (34–62) and 16 (46%) had received a previous autologous transplant. At transplant, 5 patients (14%) were in CR1 (after several lines of chemotherapy), 9 (25%) in 〉CR1, 12 (34%) in PR, 1 (3%) had stable disease (after 3 chemotherapy lines) and 8 (23%) progressive disease. All patients engrafted. Acute GVHD developed in patients 19 (54%) (17patients (48%) grade II-IV). Chronic GVHD developed in 18 out of 27 patients at risk (67%), being extensive in 11 (41%). Disease was evaluated at day +100 and at that moment 23 patients were in CR, (85%) 1 (4%) in PR, two (7%) had stable disease and 9 patients ( 26%) have died. With a median follow up of 60 months (range: 32–80 months), 20 patients (57%) are alive disease free, and 14 (43%) have died, 12 of them (37%) due to transplant toxicity and 2 patients (6%) due to progression. Overall Survival (OS) and Event Free Survival (EFS) are 57 and 54 % respectively. Analysing variables which influence on OS and EFS, patients 55 years have a OS significantly shorter than those

Description: Abstract 3219 Poster Board III-156 Introduction Dimethylsulfoxide (DMSO) containing cryoprotective solutions are routinely used for the storage of hematopoietic progenitors (HP). At room temperature, DMSO is toxic for the cells and may produce severe adverse reactions during their infusion, especially in the pediatric patients. These problems can be avoided by washing the cells prior to the infusion. Our objective was to test if an automatic washing method (Sepax S-100, Biosafe) allowed us to preserve the CD34+ cell numbers with an adequate viability and engraftment potential. Material and Methods Forty five peripheral blood HP apheresis that have been cryopreserved using autologous plasma plus 9% DMSO were studied. After rapid thawing in a water bath at 37° C, an automatic wash with the Sepax S-100 (2 washes cycle) was performed. Nucleated cell levels determined by an hematology analyzer, flow cytometry CD34+ cell counts and Trypan Blue cell viability test were performed on aliquots collected prior to and after the washing technique. The paired Student's t-test and the Pearson's correlation coefficient were used for the statistical analysis. Results The mean total nucleated cell (TNC) and CD34+ cell recovery was 75,47% ± 3, and 94,66% ± 4,62 respectively. In spite of the TNC significant loss (p500 cells /mL) and platelet engraftment (〉50.000 cells/mL) were 11 ± 0,2 and 20 ± 1,4 days respectively. Conclusions The Sepax S-100 automatic wash protocol of DMSO containing peripheral blood progenitor cells determines a good CD34+ cell recovery and preserves their viability and engraftment potential. This method avoids the DMSO infusion related adverse events and it constitutes a closed and easy to do procedure. Disclosures No relevant conflicts of interest to declare.

Description: Introduction: The addition of rituximab to induction therapy had improved the outcome of patients with primary mediastinal B cell lymphoma (PMBCL). For those patients who are primary refractory or relapse after having achieved a remission, high-dose therapy and autologous stem cell transplantation (ASCT) is considered as standard treatment. Only scanty information, however, is available regarding the role of ASCT in patients with relapsed / refractory PMBCL in the rituximab era. Moreover, the impact of pre- and post-transplant irradiation remains uncertain. The objective of the current study was to investigate the results of ASCT for PMBCL in the rituximab era, identify variables predictive for outcome, and assess the role of adjuvant radiotherapy. Patients and methods: For this retrospective study, all EBMT registered patients with PMBCL aged between 18 and 70 years who were treated with a first ASCT between 2000 and 2012 were eligible. Baseline patient, disease, and transplant data were collected from MED-A forms. Centers with potentially eligible patients were contacted to provide additional treatment and follow-up information including a written histopathology report. Statistical analysis used log rank test to assess the impact of baseline characteristics on survival endpoints. In multivariate analysis, the relevance of prognostic factors was estimated using Cox regression models. Curves of cumulative incidence of relapse (IR) were compared by Gray's test. Multivariate analysis of IR used Fine and Gray models. Results: 86 patients with confirmed PMBCL were eligible and had the full data set required for this analysis. 51% were female, median age was 34 years (range 20-69). Median time from diagnosis to ASCT was 12 months (12-299). 63.5% of the patients presented with a bulky mediastinal mass, larger than 10cm at diagnosis, 30.5% had stage IV disease, and 44% had B symptoms. 92% had received at least 2 lines of therapies, 85% had rituximab and 30% had received radiotherapy prior to transplantation. At ASCT, 11% still had a mass greater than 10 cm, and 19% a mass of 5-10cm. Remission status at ASCT was CR/PR1 in 21% of the patients, CR/PR〉1 in 51%, and refractory disease in 28%. 31 patients (41%) received irradiation post-transplant. Thirteen patients of 24 patients (54%) transplanted in PR attained CR at day +100 post ASCT. With a median follow-up of 39 months (24-73), 3-year non-relapse mortality, IR, event-free survival (EFS) and overall survival (OS) for the whole series were 9%, 33%, 58% and 71%, respectively. By univariate analysis, refractory disease at ASCT and residual mass 〉 5cm at ASCT were significant adverse predictors for IR, EFS, and OS. 3-year EFS was 35% in refractory subjects vs 66% in chemosensitive patients (p=0.001), and 100% in those autografted in CR/PR1 vs 60% in those transplanted in more advanced response p=0.018. Notably, patients transplanted with refractory disease with a history of irradiation prior to ASCT had a superior outcome compared with non-irradiated refractory patients.Multivariate analysis suggested post-transplant irradiation to be associated with a significantly reduced IR (HR=0.24; p=0.028) and improved EFS (HR=0.24; p=0.018) and OS (HR=0.21; p=0.032). Discussion: In conclusion, this analysis gives first specific evidence that ASCT can provide durable remissions in patients with relapsed / refractory PMBCL in the rituximab era. Pre or post-transplant irradiation appears to be important, though deserves further studies. Disclosures No relevant conflicts of interest to declare.

Description: Introduction Extracorporeal photopheresis (ECP) is a cell-based immunomodulatory therapy involving the separation of peripheral blood autologous mononuclear cells followed by ex-vivo administration of 8-methoxypsoralen (8-MOP) and UVA radiation before reinfusion. ECP is efficient for the treatment of multiple diseases mediated by unregulated T cell populations, such as cutaneous T cell lymphoma, autoinmune diseases or graft-versus-host disease (GVHD), the major complication after allogeneic bone marrow transplantation. Our aim in the present work was to compare the therapeutic effectiveness of 8-MOP with other two new compounds (BB01 and BB02) in a experimental murine model of GVHD. Methods Murine GVHD was induced after transplanting bone marrow cells and splenocytes from donor Balb/c mice into C57Bl6J recipients previously conditioned with a lethal dose of 10 Gy split into two doses of 5 Gy spaced 24 hours apart. To investigate the therapeutic effectiveness of ECP with either 8-MOP, BB01 or BB02, splenocytes from separate cohorts of C57Bl6J with GVHD were isolated 12 days after transplantation, incubated with the different compounds, irradiated with UVA light and injected intravenously once a week for four weeks. Survival after transplantation was monitored daily and clinical GVHD was graded using a previously described score analyzing weight loss, posture (hunching), activity, skin integrity and fur texture (Cooke et al, 1996). Mice of each group were evaluated and graded from 0 to 2 for each criterion, obtaining a clinical index by summation of the five criteria scores (maximum index=10). Results Mice treated weekly with BB02 showed a significant higher survival than those treated with 8-MOP (p=0,038), while BB01 had a similar effect to that of 8-MOP. Mice treated with either compound improved their clinical GVHD score compared to untreated mice group, being significantly lower with BB02 than with BB01 and 8-MOP (p=0,023). Conclusions BB02 was more efficient than 8-MOP in the reversal of murine GVHD, while BB01 showed the same therapeutic effectiveness than 8-MOP. Acknowledgments Work financed by the Spanish Ministry of Science and Innovation (Ref: BFU2010-19599) and the Spanish Net of Cell Therapy (TerCel) from Institute of Health Carlos III. Disclosures: No relevant conflicts of interest to declare.

Description: Introduction: Extracorporeal photopheresis (ECP) is a clinically used cell-based immunomodulatory therapy that involves exposure of peripheral blood autologous mononuclear cells to the photosensitizer 8-methoxypsoralen (8-MOP) and UVA radiation before reinfusion. ECP has demostrated efficacy in the treatment of multiple diseases caused by unregulated T cell populations such as graft-versus-host disease (GvHD). During UVA irradiation, 8-MOP covalently binds to DNA, inducing apoptosis. It has been previously reported that 8-MOP-mediated apoptosis triggers some immumodulatory effects that finally lead to the acquisition of peripheral immunotolerance, including the generation of tolerogenic dendritic cells and T regulatory cells (Treg), production of anti-inflammatory cytokines such as IL-10 and TGF-β and the deletion of effector CD4+ and CD8+ T cells in lymph nodes, mechanisms that account for the clinical benefit of this therapy. In the present work the in vitro activity and immunomodulatory effects mediated by 8-MOP in comparison with other two new photosensitizer (BB01 and BB02) has been analyzed, aiming to find novel more effective photochemotherapeutic compounds to use in ECP. Moreover, the therapeutic effectiveness of BB01 and BB02 by ECP was also evaluated in an experimental murine model of GvHD. Methods: Human mononuclear cells (MNC) were incubated with increasing concentrations of 8-MOP, BB01 and BB02 and irradiated with UVA light. The MNC apoptosis percentage was measured by flow cytometry after 48h of culture at 37ºC. Also, mixed lymphocyte cultures (MLC) with either myeloid immature (iDC) or mature dendritic cells (mDC) and MNC treated with the different compounds and UVA radiation were performed. After 48h of MLC, CCL21-stimulated migration of iDC and mDC was studied. In other assays, after 6 days of MLC the proliferation of treated MNC, production of pro-inflammatory and anti-inflammatory cytokines, generation of tolerogenic DC and differentiation of Treg were analyzed. On the other hand, murine GvHD was induced after transplanting bone marrow cells and splenocytes from donor Balb/c mice into C57Bl/6J recipients previously conditioned with a lethal dose of 10 Gy. For doing the ECP procedure, splenocytes from separate cohorts of C57Bl/6J with ongoing GvHD were isolated 12 days after transplantation, incubated with the different compounds and UVA light and injected intravenously once a week for four weeks. Survival after transplantation was monitored daily and clinical GvHD was graded using a previously described score analyzing weight loss, posture, activity, skin integrity and fur texture. Results: There was a significative increase of MNC apoptosis when usingBB02 (from 50 ng/ml, p

Description: Introduction: To discriminate different outcomes among patients in CR, the International Myeloma Working Group (IMWG )introduced more stringent CR (sCR) criteria by adding to the pre-existing CR parameters the requirement of a normal free-light chain ratio (sFLCr) plus the absence of clonal plasma cells (PCs) in bone marrow (BM) by immunohistochemistry (IHC). In 2011,the low-sensitivity cytometrycriteria were included as alternative methodology to IHC to define sCR. Aim: To validate the preliminary data of our previous study (Blood 2015. 126:858-62) regarding the lack of influence of an abnormal sFLCr in the outcome of MM patients, through the analysis of a more extensiveseries of newly diagnosed multiple myeloma (NDMM) patients in CR or sCR. Patients and Methods: This study is based on 459 NDMM patients who were transplant candidates and enrolled in the GEM2012MENOS65phase 3trial;evaluable patients were enrolled in a subsequent maintenance trial (NCT02406144).CR and sCR was defined according to the IMWG criteria. Agreeing to the protocol, patients with 10-4 MRDsensitivity). If we increase the sensitivity of the MRD to 10-6, the differences in PFS at 2 years are more evident (2 years-PFS 94% vs 67%; P10-6 sensitivity). Conclusion: These results confirm our previous findings based on GEM05menos65/ GEM10mas65 clinical trials, indicating that for MM patients stringent CR criteria does not predict a different outcome as compared to standard CR. Specifically, the sFLCr doesn't identify patients in CR at distinct risk. If this essential criterion in the definition of sCR lacks connotations for the prognosis, is it not justified to maintain a response category whose real significance depends on the combination of the traditional CR criteria with a negative MRD status based on very low (IHC) or low resolution (