ALBERT

Description: Introduction DLIs represent a major therapeutic approach for relapse and mixed chimerism (MC) after allogeneic hematopoietic cell transplant (AlloHCT). DLI studies have identified several variables with impact on response and GvHD. Despite some studies having explored the role of T-cells and other cell subsets, such as mononuclear cells (MNCs), comprehensive data regarding the cellular composition of DLI and its role in GvHD remains incomplete, as the development of GvHD post DLI is often unpredictable. Herein we analyzed the cellular composition of DLI from fully human leukocyte antigen (HLA) identical sibling (HLA Id Sib) donors and its impact on the development of GvHD in patients who underwent AlloHCT for hematological malignancy, and its impact on the development of GvHD. Methods Inclusion criteria were as follows: 1) Patients ≥ 18 years-old, 2) AlloHCT, 3) HLA Id Sib donor; 4) treatment with DLI; and 5) signed informed consent of patient and donor. Exclusion criteria were: 1) unrelated or mismatched related donors, 2) HCT2 prior to DLI, or 3) GvHD at DLI. For the purpose of avoiding bias, only the cell composition of the first DLI (DLI1) was analyzed. The following cell subsets of the DLI were studied: CD3+, CD4+, CD8+, CD16+CD56+CD3+ (NKT-cell), CD3+CD45RA+CCR7+ (TN), CD3+CD45RA+CCR7+CD31+ (TRTE), CD3+CD45RA+CD95+CD27+ (TSCM), CD3+CD45RA-CCR7+ (TCM), CD3+CD45RA-CCR7- (TEM), CD3+CD45RA+CCR7- (TTE), CD3+CD4+CD25brightCD127dim (TREG), CD3+CD4+CD25brightCD45RA+CD127dim (naïve TREG). The TN, TCM, TEM and TEM compartment was analyzed for both CD4+ and CD8+. We also analyzed the MNCs, CD19+ (B-cell), CD27+CD19+ (mature B-cell), CD16+CD56+CD27- (natural killer (NK+) cell) and CD16+CD56+CD27+ (CD27+NK+cell). Results Fifty-six DLIs were infused in 36 patients; the median number of DLI was 1 per patient (range, 1-3). Diagnoses were as follows: 13 AML/MDS, 6 HL, 5 MPN, 4 NHL, 4 CLL, 3 MM and 1 B-ALL. For the study, a landmark analysis was performed from the DLI date. The median follow up from DLI was 282 days (range, 9-5,560 days). Overall response rate in relapsed patients was 29% (9 of 31 patients; 6 CR and 3 PR, most responses being observed after DLI1. Further, five patients had DLI for MC and full donor chimerism was achieved in all patients. Thirteen patients (36%) developed GvHD post DLI. Two patients had GvHD before DLI, but there was no case of GvHD at DLI. The median time interval form DLI to GvHD was 76 days (range, 7-261). As per clinical presentation, 10 patients (27%) had acute GvHD, whereas eight patients (22%) had chronic GvHD. The 6-month and 1-year cumulative incidence (CI) of GvHD was 33% and 46%, respectively. When the risk of GvHD was analyzed according to DLI cell subsets, we observed that a DLI1 containing 〉3x106 CD8+TN correlated with an increased incidence of GvHD (Figure 1a). Also, a DLI1 with 〉0.8x108 MNCs/Kg (Figure 1b), 〉2.6x106 mature B-cell/Kg, or 〉0.35x106 CD27+NK+cells/Kg were linked to the development of GvHD (Table 1). Noteworthy, CD3+, TN (both CD4+ and CD8+ combined) or CD4+TN had no impact on the development of GvHD; and a high proportion of TREG was not protective for the development of GvHD (Table 2). Finally, there was no statistically significant association between any clinical variable and GvHD. Conclusion In conclusion, in this cohort of AlloHCT patients from HLA Id Sib donors, a DLI1 containing a high proportion of CD8+TN, but not CD4+TN, increased the probability of developing GvHD. Further, a DLI1 containing a high dose of MNCs, CD27+NK+cells and mature B-cell also associated with GvHD. These data provide novel insight for the understanding of GvHD post DLI. A DLI1 containing a lower dose of CD8+TN could reduce the risk of GvHD, but this asset warrants further validation in larger cohorts, and within a controlled randomized trial setting. Disclosures Bosch: F. Hoffmann-La Roche Ltd/Genentech, Inc.: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Kyte: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Celgene: Honoraria, Research Funding; Acerta: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Takeda: Honoraria, Research Funding; AstraZeneca: Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Description: Abstract 3375 Poster Board III-263 Reduced-intensity conditioning (RIC) allogeneic hematopoietic cell transplantation (Allo-HCT) has become a feasible and effective therapeutic approach for younger patients with relapsed or refractory follicular lymphoma (FL). However, there is still much debate regarding the most appropriate conditioning regimen or whether the use of in vivo T-cell depletion (TCD) is beneficial or not for these patients. We analyzed the outcome of 164 patients with advanced FL reported to the EBMT from 1999 to 2007, who underwent RIC Allo-HCT conditioned with fludarabine plus an alkylating agent. Donors were HLA-matched siblings in all cases. Patients receiving transplants from alternative donors or conditioned with other agents were specifically excluded. The alkylating agent was melfalan in 48% of cases, busulfan in 32% and cyclophosphamide in 20%. Forty-six patients (28%) received anti-thymocyte globulin (ATG), 41 (25%) received alemtuzumab and 77 (47%) did not receive TCD in vivo. Median age at transplantation was 50 (range 29-64) years, and patients receiving alemtuzumab were significantly younger [45 (33-63)] than those receiving ATG [52 (29-64)] or no TCD [50 (32-64)], P = 0.05. There were no other differences among groups in terms of disease stage or presence of bulky masses at diagnosis, interval from diagnosis to HCT, number of prior therapies, or disease status at HCT. Engraftment was observed in 161 (98%) patients, with no significant differences among groups. Median follow-up was 43 (1–110) months for survivors. At three years, non-relapse mortality (NRM), relapse rate (RR), progression-free survival (PFS) and overall survival (OS) were 17% (95% CI 12-24%), 23% (17-31%), 60% (52-68%) and 75% (67-82%), respectively, for the entire cohort. The incidence of grade 2-4 acute graft-versus-host disease (GVHD) was significantly higher for patients not receiving any TCD (31%) compared to TCD patients (18%), P = 0.05, and the incidence of chronic GVHD at one year was also significantly higher for the former compared to the latter group (68% vs. 25%, P 〈 0.001). There were no significant differences in NRM among groups, but there was a trend towards a higher RR in patients receiving alemtuzumab (40%) or ATG (24%) compared to patients receiving no TCD (16%) (P = 0.15), which translated into a trend towards a significantly shorter 3-year PFS for the alemtuzumab group (42% vs. 69%; P = 0.18). However, there were no differences in the 3-year OS among groups, which was 77% for patients receiving alemtuzumab, 73% for those receiving ATG and 77% for patients not receiving any TCD. In conclusion, results with RIC Allo-HCT from HLA-identical siblings were very promising for patients with advanced FL. Both alemtuzumab and ATG were effective in reducing acute and chronic GVHD, but had no significant impact on NRM. There was a trend towards a shorter PFS for patients receiving alemtuzumab, which did not translate into a significantly different OS. Disclosures: Delgado: Bayer Schering Pharma: Consultancy, Research Funding; Genzyme: Research Funding. Off Label Use: The use of alemtuzumab as a T-cell depleting agent in the context of hematopoietic transplantation is considered off-label.

Description: The outcome of patients (pts) with PTCL receiving conventional therapy is dismal. Because of this, there is an increasing interest to investigate intensive treatments in these patients. The aim of this study was to analyze the toxicity, response and outcome of a phase II trial that includes high-dose chemotherapy (CT) plus ASCT as first-line treatment for pts with PTCL. Forty-one pts (30M/11F; median age: 47 yrs.) diagnosed with PTCL (excluding cutaneous and anaplastic ALK+), in stages II-IV and 0.1). Four-year OS was 57% (95%CI: 31–83%) and 71% (95%CI: 37–100%), respectively (p〉0.1). In summary, in this series of patients with PTCL a relatively high CR rate was obtained with high-dose CHOP/ESHAP followed by ASCT. Toxicity was manageable. The contribution of ASCT to pts outcome is debatable because of the absence of significant differences in OS and EFS of patients in CR transplanted vs. those not transplanted. Novel strategies aimed at increasing the CR rate in these patients warrant investigation.

Description: Although nonmyeloablative conditioning regimen transplantations (NMTs) induce engraftment of allogeneic stem cells with a low spectrum of toxicity, graft-versus-host disease (GVHD) remains a significant cause of morbidity and mortality. In vivo T-cell depletion, using alemtuzumab, has been shown to reduce the incidence of GVHD. However, this type of maneuver, although reducing GVHD, may have an adverse impact on disease response, because NMTs exhibit their antitumor activity by relying on a graft-versus-malignancy effect. To explore the efficacy of alemtuzumab compared with methotrexate (MTX) for GVHD prophylaxis, we have compared the results in 129 recipients of a sibling NMT enrolled in 2 prospective studies for chronic lymphoproliferative disorders. Both NMTs were based on the same combination of fludarabine and melphalan, but the United Kingdom regimen (group A) used cyclosporin A plus alemtuzumab, whereas the Spanish regimen (group B) used cyclosporin A plus MTX for GVHD prophylaxis. Patients receiving alemtuzumab had a higher incidence of cytomegalovirus (CMV) reactivation (85% versus 24%,P 

Description: Abstract 650 The Seattle group previously reported that the adapted Charlson CI (Sorror-CI) was useful for predicting NRM and survival in patients undergoing allo-SCT. However, the value of this CI index is still under considerable debate, since the cohort used by Sorror et al. (Blood 2005; 106:2912-9) included a large age range, a wide variety of diseases, with both myeloablative and RIC transplants. On the other hand, toxicities and NRM prediction is most likely needed in elderly and medically infirm patients who are increasingly offered RIC allo-SCT. Therefore, the current study was designed to test the performance of this CI and its association with outcomes among a cohort of 345 patients (i) aged 〉50 y.; (ii) diagnosed with a single disease entity, AML in CR1, and (iii) who underwent a RIC allo-SCT reported to the EBMT registry between 1999 and 2006. In this series, the median year of allo-SCT was 2004, and the median age was 58 y. (range, 50-76). 32% of patients needed more than one induction course to achieve CR1. A fludarabine-based RIC regimen was used in 64% of patients, while 31% of patients received low-dose TBI as part of their RIC, and 6% received other non-specified RIC regimens. 76% of the patients received allo-SCT from an HLA-matched sibling donor. Based on score calculated with hazard ratios (HR) estimated on the population studied, 161 patients (47%) had a CI score of 0, 96 patients (29%) had a score of 1, and 49 (14%) had a CI score of 2. The remaining 39 patients (11%) had CI scores of 3 or more. In this cohort, 2 years overall and leukemia-free survival rates were 64±3% and 54±3% and the 2 years relapse and -NRM cumulative incidences were 32±2%, and 15±2% respectively. The 2 years NRM incidences according to comorbidities score 0, 1, 2 and 3+ were 9±2%, 15±4%, 18±5% and 31±7%, respectively. In multivariate models (adjusted for recipient age, donor type, use of TBI or not, and cytogenetics risk group) comorbidities such as moderate active liver disease, obesity, prior history of renal dysfunction, and prior history of severe liver disease were associated with the highest HRs for 2 years NRM (varying from 2.11 to 2.76) and 2 years cumulative incidences of NRM varying from 22% to 44%, whereas previous solid tumor, diabetes, rheumatologic abnormalities, moderate pulmonary diseases, cardiac abnormalities (other than arrhythmia and valve disease) were associated with the lowest HRs (varying from 0.2 to 1.0) with 2 years cumulative incidences of NRM varying from 5 to 17%. Results from this large study performed in a single disease entity and homogeneous allo-SCT setting, suggest that the hematopoietic cell transplantation-specific CI is a simple, informative and useful tool for capturing pre-transplant comorbidities, and for predicting NRM after RIC allo-SCT for AML in CR1 in patients aged 〉50 y. Such index may be used for clinical trials and patient counselling before RIC allo-SCT. Disclosures: No relevant conflicts of interest to declare.

Description: Peripheral blood (PB) has become the major source of hematopoietic stem cells for autologous stem cell transplantation (ASCT) in the last 15–20 years. Nevertheless, there is a subset of patients who do not mobilize adequate numbers of CD34+ cells. There are no clearly established guidelines with respect to second-line mobilization protocols. The aim of this study has been to analyze our experience as a single center with this population of poor mobilizers trying to identify clinical or biological adverse prognostic factors associated to a poor mobilization of progenitor cells into PB, results of second- and third-line mobilization procedures and outcome after the ASCT of those patients who could be autografted in terms of hematological recovery. Poor mobilizing patients were defined as those in whom the apheresis procedure could not be started because of 〈 10 CD34+ cells/ul or those in which a less than 2 × 106 CD34+ cells/kg could be collected in the first mobilization attempt. From January/2000 to January/2008, 126 patients [70 males/56 females, median age of 53 years (range, 20–70)] out of a total number of 450 patients mobilized for an ASCT in our institution (28%) were identified as poor mobilizers. Clinical diagnosis were: 29 multiple myeloma, 16 Hodgkin’s lymphoma, 48 non-Hodgkin’s lymphoma, 28 acute leukemias and 5 other diagnosis. Median time from diagnosis to mobilization therapy was 19 (range, 3–120) months and median number of chemotherapy lines received before the procedure was 2 (range, 0–5). The first mobilizing protocol was G-CSF alone (5–10 ug/kg/day sc) in 72% of the patients or the combination of chemotherapy plus G-CSF in 28% of the patients. A second mobilization procedure was attempted in 34 patients (28%) with high-doses of G-CSF alone (16–20 ug/kg/day sc) in 24 patients, the combination of G-CSF plus chemotherapy in 8 patients and the combination of G-CSG with stem cell factor (SCF) in 2 patients. A third mobilization attempt was performed in 6 patients (high-doses of G-CSF alone in 4 patients, G-CSF plus chemotherapy in 1 patient and G-CSF plus SCF in 1 patient). Sixty-nine patients (54%) were finally autografted. Median number of CD34+ cells/kg infused were 2.15 × 106/kg (range, 1.01–4.00). Median time to neutrophil recovery after transplantation was 11 days (range, 4–20). Patients with an inadequate mobilization constitute a significant clinical problem (25% of the whole population of patients with an indication of ASCT in our centre). Nevertheless, half of these patients can be rescued for an ASCT procedure with one or two more attempts. Neutrophil recovery after the autologous transplant in those patients undergoing the procedure seems to be similar to that of the group of patients with an adequate first mobilization attempt. New mobilizing agents should be investigated in order to increase the efficacy of the mobilization processes.

Description: The outcome of patients (pts) with PTCL receiving conventional therapy is dismal. Because of this, there is an increasing interest to investigate intensive treatments in these pts. The aim of the study is to analyze the interim results, in terms of toxicity, response and outcome, of a phase II trial that includes high-dose chemotherapy (CT) plus ASCT as 1st-line treatment for pts with PTCL. Thirty-four pts (23M/11F; median age: 47 yrs.) diagnosed with PTCL (excluding cutaneous and anaplastic ALK+), in stages II–IV and ≤65 yrs, who have already finished planned therapy, are the subject of this analysis. Nine pts (26%) presented with primary extranodal disease, 24 (71%) were in stage IV, and 13 (38%) had bone marrow involvement. Fifty percent of the pts had high/intermediate or high-risk IPI, whereas 53% were in the groups 3 or 4 according to the Italian Index for PTCL. Pts received intensive CT (3 courses of high-dose CHOP [cyclophosphamide 2000 mg/m2 day 1, adriamycin 90 mg/m2 day 1, vincristine 2 mg day 1, prednisone 60 mg/m2/day, days 1 to 5, with mesnum and G-CSF], alternating with 3 courses of standard ESHAP). Responders (CR or PR) were submitted to ASCT. Median follow-up of surviving pts was 3.9 yrs (range, 0.5–7.6). Twenty four pts (71%) received the planned 6 courses of CT. Response rate after CT was: CR, 12 cases (35%); PR, 8 (23%); failure, 14 (42%), including one pt who died because of sepsis. Hematological toxicity of CT mainly consisted of neutropenia (grades 3–4 in 87 and 62% after high-dose CHOP and ESHAP, respectively) and thrombocytopenia (grades 3–4 in 63 and 68%, respectively). Severe infection requiring hospitalization was observed in 38 and 15% of courses of high-dose CHOP or ESHAP, respectively. Only 14 of the 20 candidates (70% of all candidates and 41% of all pts) received ASCT due to lack of stem-cell mobilization (3 cases), previous toxicity (2) and pt decision (1). No differences in the outcome were seen among these 20 pts according to whether or not they could eventually receive ASCT. No major toxicity was observed after ASCT. The overall response after the whole treatment was: CR, 14 cases (41%), PR, 5 (15%), failure, 15 (44%). Two of 14 pts in CR and the 5 pts in PR eventually progressed. Four-year failure-free survival (FFS) was 30%, whereas 4-year disease-free survival for pts achieving CR was 63%. Nineteen pts have died during follow-up, with a 4-yr overall survival (OS) of 38% (95%CI: 21–55%). Most patients died because of PTCL progression, but 2 pts died in CR due to a secondary acute lymphoblastic leukemia and to a lung cancer, respectively. IPI and Italian Index for PTCL were able to predict FFS and OS. In summary, in this series of patients with PTCL a relatively high CR rate was obtained with high-dose CHOP/ESHAP followed by ASCT. Toxicity was manageable. However, the prognosis of patients with PTCL, particularly of those not achieving CR, is still very unfavorable. Figure Figure

Description: BACKGROUND: Systemic fungal infections including invasive candidiasis and aspergillosis are a significant cause of mortality and morbidity in immunocompromised patients. ABLC is used for the treatment of fungal infections in patients who are intolerant or unresponsive to conventional amphotericin therapy. Caspofungin is a second-line treatment for invasive aspergillosis. The different mechanisms of action of these agents support a dual-therapy regimen; synergistic results have been observed in vitro (Arikan et al, Antimicrob Agents Chemother, 2002;46:245–47). The aim of this open-label, multi-centre study was to investigate safety, tolerability (primary endpoints) and efficacy of concomitant ABLC and caspofungin in immunocompromised patients with refractory FUO (fever of unknown origin). METHODS: At baseline patients were required to have serum creatinine≤220 μm/L, neutrophil count

Description: Background: Pneumococcal infections are causes of death after SCT. The IDWP01 trial compared early (E) vs late (L) pneumococcal immunization after allogeneic SCT, starting either at 3, or at 9 months post-transplant with 3 doses of PCV7 given at 1 month intervals,. This study has shown that the early immunization was not inferior in the proportion of responders one month after the last PCV7 dose. In addition, all patients received one dose of PPV23 after the 3 doses of PCV7 at 12 months and 18 months in the E and L groups, respectively. The goal of this supplementary analysis was to look at the immune response to the PPV23 according to the time of its administration. Methods: 158 patients were randomized in the IDWP01 trial, 75 in the E, and 83 in the L group, of whom 109 received the PPV23 (56 in the E, and 53 in the L group, respectively) and 86 (44 in the E, and 42 in the L group) were followed up to 24 months after SCT. The immune response to PPV23 was assessed by the antibody (Ab) titres, measured by ELISA to pn1 and pn5, which are both included in PPV23 but not in PCV7. Pn1 and pn5 Ab titres were assessed before, and one month after the PPV23 dose, and at 24 months post-SCT. Two cut-off levels for protection were considered for each antigen: □ .15 μg/ml and □ .5 μg/ml. The geometric mean concentrations (GMC) were also analyzed. Additionally, all Ab titres to the 7 PCV7 antigens were measured in parallel. Results: Ab titres to pn1 and pn5 before the PPV23 were not significantly different in the E (at 12 mo post-SCT) and in the L (at 18 mo post-SCT) group. Whatever the cut-off used to evaluating the response, the percentage of responders one month after the PPV23 was not significantly different in both groups: For pn1: □ .15μg/ml: E: 80% vs L: 87%; □ .5μg/ml: E: 42% vs L: 60%; For pn5: □ .15μg/ml: E: 84% vs L: 94%; □ .5μg/ml: E: 62% vs L: 74%). The GMC were not significantly different in the E vs. the L group for pn1. The GMC were significantly higher in the L vs. the E group for pn5 (P=.02). At 24 months post-SCT, there was no significant difference in the GMC of pn1 and pn5 Ab titres but there was a tendency for higher titres in the L group for both antigens. Additionally, among 36 non-responders to the PCV7 at the time of PPV23 administration, 21 remained non-responders, but 15 (42%) responded to all the seven PCV7 antigens 1 month later. Conclusion: Although there was a tendency for a better response rate and for higher GMC for pn1 and pn5 according to the timing of immunization with PPV23 (12 vs. 18 months) in favour of the L group, these differences were not significant. Additionally, PPV23 increase the response rate to the serotypes included in the PCV7. Therefore, we recommend starting PCV7 vaccination early after SCT (3 PCV7 from 3 months). A dose of PPV23 after 3 doses of PCV7 broadens the immune response to pneumococci as well as increases the response rate to the serotypes included in the PCV7 and should therefore be considered at 12 months after SCT. The authors are grateful to the Safety committee: D Engelhard, P Reusser, and P Reinert