ALBERT

Description: Invasive fungal infections are a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation(allo-HSCT). During the past 2 decades, changes in transplantation practices and strategies to diagnose and treat IFI have likely impacted the epidemiology of IFI. Polymorphism of genes, which take part in the allogeneic immune response after allo-HSCT, lead to allograft immunoreactivity differences between individuals. It also may affect IFI after transplantation. Methods: We analyzed allo-HSCT recipients at our center during the period January 2001 through March 2009. We analysis gene polymorphism of innate immune gene TLRs among 240 pairs of specimens of recipients and donors, in order to find out IFI individual difference and high risk genotype of IFI after transplantation. Results: There were 99 patients (41.2%) occured IFI after allo-HSCT. Hematological malignant diseases, serious acute GVHD, and extensive chronic GVHD were risk factor for IFI after allo-HSCT. Univariate analysis of the relationship between risk factors of IFI after all0-HSCT and the TLRs gene polymorphism, we found that in the 10 SNP loci in 5 TLRs , two polymorphisms of TLR8 gene, (+1 A/G, rs3764880; +354 C/T, rs2159377) effect the incidence of IFI after allo-HSCT. Other gene polymorphism of TLRs (TLR1, TLR2, TLR3, TLR9) had no significant effects on the risk of IFI after allo-HSCT. Conclusion: The study of the relationship between the gene polymorphism of natural immune molecule TLRs and the risk factors after allo-HSCT showed that TLR8 genotype of donor stem cell significantly influenced the incidence of IFI after allo-HSCT at the first time. It provide the evidence to establishment a fungal infection index system, which based on risk stratification of genetic background before transplantation of donors and recipients stem cell, and provide basis for selection of unrelated donor and prevention and treatment of IFI Disclosures No relevant conflicts of interest to declare.

Description: Abstract 4546 Hemorrhagic cystitis (HC), occurring mostly within first month after allogeneic hematopoietic stem cell transplantation (allo-HSCT), is a common complication which often requires prolonged hospitalization, considerable expense, and occasionally causes significant morbidity. The data from different transplantation centers about this complication are different in the incidence, risk factors, therapeatic strategies and outcome. In this study, we retrospectively analyzed the incidence, risk factors, prophylaxis and treatment regimens of HC after unrelated donor HSCT(URD-HSCT)in our center From October 2000 to February 2008, 168 consecutive patients underwent URD-HSCT were enrolled,including 113 male patients and 55 female patients. The median age was 26 years (range,8-52 years). The main myeloablative conditioning regimens used were busulfan/cyclophosphamide (BuCy) without total body irradiation (TBI); Reduced-intensity conditioning regimens (RIC) were predominantly fludarabine-based combinations without irradiation. Anti-thymocyte globulin(ATG) were added to the conditioning regimen in the patients receiving HLA-mismatched URD-HSCT. The prophylaxis regimen for HC included intravenous mesna,hyperhydration and intermittent diuretic. The diagnosis of HC was based on the presence of sustained microscopic or macroscopic hematuria in the absence of other clinical conditions such as urinary bacteria and fungi infection. It could be clinically graded to I to IV according to hematuria and divided into early onset (EOHC) and delayed (LOHC) according to the time of onset. In total, 27 of 168 patients (16.1%) developed HC at a median interval of 40 days post engraftment (range, 8–89 days). Among them,11 patients developed grade I (6.5%), 10 patients developed grade II (6.0%), 6 patients developed grade III (3.6%), while no patients with grade IV. Five patients(2.98%) presented EOHC and 22 patients (13.17%) presented LOHC. We discovered that in 27 patients with HC only 3 were female (11.1%), however in 141 patients without HC, 52(36.9%) were female (P

Description: Key Points We developed an approach of T-cell-replete haploidentical HSCT with low-dose anti-T-lymphocyte globulin. Outcomes of suitably matched URD-HSCT and HRD-HSCT are similar, and HRD-HSCT improves outcomes of patients with high-risk leukemia.

Description: Acute graft-versus-host disease (GVHD) remains the major barrier of allogeneic bone marrow transplantation (allo-BMT). In the current view, GVHD fundamentally depends on donor T cells interaction with host derived dendritic cells (DC) leading to their activation, proliferation, and differentiation. We and others have demonstrated that transforming growth factor β1 (TGF-β1) treated DC (TGFβ-DC) have regulatory characteristic and could induce allogeneic specific immune tolerance in vitro. In the current study, we focused on the effects of recipient-derived TGFβ-DC in a murine GVHD model. After total body irradiation, 5.0×106 recipient-derived TGFβ-DC were injected into C57BL/6 (H-2b) with bone marrow-splenocyte grafts from major histocompatibility complex (MHC) disparate BALB/c mice (H-2d). Survival analysis showed TGFβ-DC co-transplantation resulted in significant prolongation of allograft survival and mean survival time (MST) was 44.3±4.5 days (p

Description: Allogeneic hematopoietic stem-cell transplantation (Allo-SCT) has become the standard of care for a number of malignant and non-malignant disorders. Neurological complications associated with these procedures remain frequent. These complications result from a variety of causes. Known risk factors include the underlying disorder, therapies used for immunosuppression, especially the use of cyclosporine A or FK506, and toxicities from radiation or chemotherapy, but other risk factors might also be of importance. We retrospectively analyzed neurological complications of 152 patients (range 10–49 years old) who underwent Allo-SCT in our transplantation center between August 2005 and July 2008. In all, there were 95 (62.5%) cases of unrelated donor transplantation, 54 (35.5%) cases of sibling donor transplantation, and 3 (2.0%) cases of haploidentical transplantation. Post-SCT neurological complications were seen in 16(10.5%) patients. They were seen in 4 cases after related donor SCT and in 12 cases after unrelated donor SCT. Neurological symptoms occurred between day −4 day and +512 day after transplantation. The complications included 4 cases with relapsed leukemia of central nervous system (2 in ALL, 1 in CML, 1 in AML), 3 cases with encephalorrhagia, 3 cases with epilepsy, 2 cases with demyelinating disease, 2 cases with encephalitis, 1 cases with multiple myositis,1 cases with peripheral neuritis. Seven cases died of neurological complication, including 4 with relapsed leukemia of central nervous system and 3 with encephalorrhagia. Other patients were cured. High dose gamma globulin conbined with Methylprednisolone were used in demyelinating disease. Antiepileptic agents were used in epilepsy. Cyanocobalamin was used in peripheral neuritis. In conclusion, neurological complications commonly occurred in Allo-SCT, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nervous system leukemia and demyelinating disease, multiple myositis should have clinical interests.

Description: Objective Because of the central role of the transcription factor nuclear factor-κB (NF-κB) in cell survival and proliferation in many kinds of cancer cells, NF-κB inhibitor may have a potential role in the therapy of cancer. Human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Some data report that CMV infection has a very close relation with NF-κB activation. But it is unknown what effect of NF-κB inhibitor pyrrolidinedithiocarbamate (PTDC) on the infection and activation of cytomegalovirus in mesenchymal stem cells(MSCs). Methods MSCs were infected by 1 TCID50 of CMV combined with/without 1μmol/L of PTDC. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene and GAPDH gene. Flow Cytometry was used to detect the CMV PP65 antigen positive cells. Results Shape of some MSCs changed after the infection of 1 TCID50 of CMV. But in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC, Cell shape changed more dominantly, almost all cells changed from thin shuttle shape to round and thick ball shape. While in the cells treated only with PDTC, shape of the cells did not changed. RT-PCR assay showed that there was a very bright band of CMV IE mRNA in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC compared with the cells infected only with 1 TCID50 of CMV. More CMV pp65 antigen positive cells were found in MSCs infected by 1 TCID50 of CMV combined with 1μmol/L of PTDC with Flow Cytometry. Conclusion NF-κB activation may affect CMV infection of cells. NF-κB inhibitor PTDC can increase the infection and activation of CMV in MSCs and we should pay more attention to CMV infection when we use NF-κB inhibitor in clinical work..

Description: Abstract 4511 Background: Hematologic relapse after Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) is associated with a dismal prognosis. Increasing minimal residual disease (MRD) after HSCT had been proved highly effective prognostic factor for post-treatment leukemia relapse. Savage chemotherapy or intensive conditioning followed by a second HSCT may be applied, but associated with a high mortality and a low rate of complete remission. Donor lymphocyte infusion (DLI) has been shown to exert a graft-versus-leukemia (GVL) effect and has been successfully used in patients who relapsed after HSCT. Objective: In this retrospective study, we sought to gain insights of the effect of DLI on clinical outcomes such as graft versus host disease (GVHD), overall survival (OS), disease free survival (DFS), and treatment-related mortality (TRM) in patients with either relapsed hematological malignancies or increasing MRD who underwent HSCT. Methods: DLIs were administered to 25 patients [10(40%) acute myeloid leukemia, 10 (40%) acute lymphoblastic leukemia, 5 (20%) chronic myeloid leukemia]. Patients with Acute leukemia had been treated with myeloablative conditioning (BuCy). Patients with chronic myeloid leukemia had been treated with non-myeloablative conditioning(Flu,ATG and Bu). Infusion of allogeneic hematopoietic stem cells are performed at our institution from 2005 to 2010. GVHD prophylaxis consisted of CsA, MMF and MTX.10 patients were diagnosed of hematologic relapse a median of 259 (30–850) days after HSCT. 15 patients had persistent increasing MRD(monitoring with flow cytometry or RT-PCR for Fusion Gene) after HSCT. DLIs were given to patients who had relapsed hematological malignancies or persistent increasing MRD. Patients with relapsed malignancies were performed reinduction chemotherapy simultaneously. A total of 57 DLIs were administered to 25 patients a median of 809 (92-1981) days after HSCT. The median transplant dose of CD3(+) cells is 4.12*10e7/kg (2.68*10e7/kg–7.97*10e7/kg). Results: The overall response rate of DLI was 80% for CML, 90% for ALL and 80% for AML.The response rate was 93.3% in patients with increasing MRD, whereas 60% in patients with relapsed malignancies. A total of 10 patients (40%,7 with increasing MRD, 3 with relapsed malignancies) developed acute GVHD. A complete response was achieved in 72% of the patients. TRM was 12% (3 patients with increasing MRD). The Kaplan-Meier estimate of OS and DFS at 3 years after DLI were 62.5% and 60.9%, respectively, with a median follow-up of 411 (32-1509) days for survivors. In patients with increasing MRD, the 3-year OS and DFS after DLI were 78.6% and 77.8%, respectively, with a median follow-up of 531(40-1509) days. In patients with relapsed malignancies,the 3-year OS and DFS after DLI were 40% and 36.8%, respectively, with a median follow-up of 469 (32-1393) days. Conclusion: Leukemia relapse is a serious therapeutic challenge following HSCT. In this retrospective study, DLIs was proved to be an effectively therapy to prevent relapse and get a better Clinical outcome in leukemia patients. Patients undergone DLI without leukemia relapse had a better outcome than the relapse group. Further strategies are required to detect early relapse in HSCT patients, and DLI may be a strategy to prevent relapse in high risk patients. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 5036 Multiple myeloma (MM) is a malignant neoplasm of plasma. With conventional chemotherapy, the rates of complete remission (CR) or very good partial remission (VGPR) are still low. Little has been reported on Bortezomib-based therapies specifically in the Chinese pateitns with MM. Here we report our results with combination therapy based on bortezomib in the Chinese population. We investigated the efficacy and safety of Bortezomib-based therapies in previously untreated MM patients. Methods: Between June 2006 and June 2010, 61 consecutive newly-diagnosed patients with symptomatic MM were treated with combination therapies based on Bortezomib. Forty-two patients were male and 19 were female. Median age was 59 years (range 37–86 years). Forty-four patients were stage 3 according to the International Staging System, 6 patients were stage 2 and 11 patients were stage 1. The conbinations included dexamethasone, dexamethasone plus subsequent thalidomide and dexamethasone plus cyclophosphamide. In detail, Bortezomib was at the dose of 1.3 mg per square meter IV on days 1, 4, 8, 11 and dexamethasone at 20 mg per square meter IV daily on the day of bortezomib and the day after, with or without daily oral thalidomide that was escalated from 100 mg to 200 mg (BD group or BDT group) or plus cyclophosphamide at 0.2 per square meter IV on days 1 to days 4 (BDC group). Thirty-four patients were in BDT group, 12 in BD group and 15 in BDC group. All patients received a median of three cycles of therapy (range 1–6). The IMWG criteria were used for response evaluation and toxicities were evluated according to the NCI Common Toxicity Criteria version 3. Results: The proportions of patients with very good partial response (VGPR) or better were 38% (13/34), 25% (3/12) and 60% (9/15) in BDT, BD and BDC group, respectively; 44% (15/34), 33% (4/12) and 33% (5/15) achieved partial response (PR). Therefore the overall response (VGPR plus PR) were 82% (28/34), 58% (7/12) and 93% (14/15). Three patients died with severe infection without disease progression. Grade 3–4 toxicities included fatigue (4/34, 1/12 and 4/15), thrombocytopenia (8/34, 3/12 and 5/15), diarrhea (4/34, 2/12 and 2/15) and infection (7/34,3/12,6/15) in BDT, BD and BDC group, respectively. Grade 1–2 neuropathy were occurred in 20 patients (59%), 6 patients (50%) and 9 patients (60%) and grade 3–4 were occurred in 6 (18%), 1 (8%) and 1 (7%) in BDT, BD and BDC group, respectively. Herpes zoster occurred in 6 patients (18%), 1 patients (8%) and 2 patients (13%) respectively. Routine anticoagulation or anti-thrombsis were not used. Only 1 patient suffered from DVT/PE but did well with treatment. Conclusions: Our preliminary experience in Chinese patients indicated that combination chemotherapy based on Bortezomib is highly effective in newly-diagnosed multiple myeloma and BDC or BDT regimens may be more superior than BD in Chinese population. There were relative lower rates of grade 3–4 neuropathy and DVT/PE in the Chinese patients with MM receved combination chemotherapy based on bortezomib. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 4513 Background: Up to date, the knowledge concerning the predisposing factors, pathogenesis and mechanisms of isolated extramedullary relapse (EMR) of acute leuekmia (AL) has not been well elucidated. Method: We retrospectively analyzed 215 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) at our center from 1998 to 2006. All of them were adult AL patients (110 acute myeloid leukemia (AML), 102 acute lymphocytic leukemia (ALL) and 3 acute mixed lineage leukemia). 10 cases (4.7%) experienced relapse of extramedullary sites. Among these 10 patients, there were four male and six female patients with a median age of 25 years (range, 22–38 years). Six patients were AML and four patients were ALL. The median interval from diagnosis to transplantation was 9 months (range, 6–15 months). Five patients had cytogenetics or molecular abnormalities. All the patients were in first CR. All the donors were HLA-identical matched (5 siblings and 5 unrelated donor). Conditioning regimen included BU and CY without total body irradiation (TBI). Graft-versus-host disease (GVHD) prophylaxis is consisted of cyclosporin A, a short-term methotrexate and mycophenolate mofetil. Result: Ten patients experienced isolated EMR at a median of 10 months (range, 3–28months) after allo-HSCT. Sites of EMR varied widely and included CNS (leptomeninges, brain, retro-orbital tissue and paraspinal tissue), skin, bone, pelvis and breasts. Five patients experienced two or more sites of EMR. The most frequently involved in relapse sites were CNS (4 patients) and skin (3 patients). Three patients subsequently developed bone marrow relapse, two patients in 2 and 3 months after the initial EMR and the third patient in 3 months after the second onset of EMR. The remaining seven patients did not develop bone marrow involvement at a follow-up ranged from 3 to 8 months after the initial EMR. Three patients received donor lymphocytes infusion (DLI) combined with chemotherapy and (or) irradiation, two patients achieved CR and remain free of disease for 25 and 32 months after transplant whereas one died due to the second EMR and the following marrow relapse. Two patients received local surgery in combination with radiotherapy or chemotherapy and both remain free of disease for 7 and 18 months post transplant. Among five patients who were only treated with local irradiation or chemotherapy or supportive care, four died from progressive disease. Conclusion: The prognosis of EMR is generally considered to be unfavorable. An intensified regimen combined of local interventions with systemic chemotherapy may be benefit for patients. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 4368 De novo Philadelphia chromosome-positive acute myeloid leukemia is a rare condition with a poor prognosis. Allogeneic stem cell transplantation (allo-SCT) is always recommended to treat this disease although its results are not always satisfactory even in young patients. Imatinib is a protein tyrosine kinase inhibitor that has now been shown to be active in Ph+ chronic myeloid leukemia and Ph+ acute lymphoblastic leukemia, however its role in Ph+ AML has not been extensively investigated. We present two patients with de novo Ph+ AML who received Imatinib combined with chemotherapy and allogeneic stem cell transplantation (allo-SCT), followed by Imatinib maintainance treatment after allo-SCT. These patients achieved long-term disease-free survival for 34 and 44 months respectively. Patient 1: A 19-year-old woman was admitted in October 2007 with fever and fatigue for one week. Physical examination showed pallor and severe sternal tenderness. Blood routine showed WBC 93.7×109/L, Hb 80.7g/L, platelets 42×109/L, 60% blast cells. Flow cytometry analysis of marrow mononuclear cells showed 84.41% blast cells positive for CD34, 80.49% for HLA-DR, 34.63% for CD13, and 5.71% for MPO. Karyotype at diagnosis was 46, XX, t (9; 22) (q34; 11) in 10 analyzed metaphases. Major BCR/ABL rearrangement (M-BCR-ABL, P210) was positive tested by real-time quantitative polymerase chain reaction(RQ-PCR). She was diagnosed with Ph+ AML (FAB M0). The patient was treated with 2 courses of DA chemotherapy then achieved bone marrow remission after the first course and achieved Complete Hemotology Response (CHR) after the second course. Then another 2 courses of high dose cytarabine were given. The status of minimal residual disease (MRD) was monitored by RQ-PCR for BCR-ABL/ABL ratio. After that BCR-ABL/ABL ratio was 68%, and then oral treatment with Imatinib 300mg/d×14d was started 4 months after diagnosis. Then 2 more cycles of Imatinib (300 mg/day for 14 days) following IA consolidation chemotherapy were taken and the patient maintained CHR and achieved statuses Complete Molecular Response (CMR) by her BCR-ABL/ABL ratio reducing to 0.4% 8 months after diagnosis. Then she took another course of Imatinib 400mg×4w therapy before allo-SCT. The patient underwent allo-SCT from a HLA-identical unrelated donor 9 months after diagnosis. STR analysis showed completely donor phenotype 1, 3 and 6 months after SCT. Imatinib (300 mg/day) was continued 74 days after SCT and the patient remains continuous CHR with Complete Cytogenetic Response (CCyR) and CMR till now. Patient 2: A 46-year-old man was admitted in Dec. 2004 with fever, cough and fatigue. Physical examination showed pallor and a small submaxillary lymphnode on the right. Blood Routine showed Hb 59g/L, platelets 115×109/L, WBC 3.1×109/L, and 24% blast cells. Bone barrow immunophenotype was CD34+, CD13+, CD117+, HLA-DR+ and MPO-. Karyotype analysis showed 46, XY, t (9; 22) (q34; 11) in 10 analyzed metaphases. Major BCR-ABL rearrangement was positive. He was diagnosed as Ph+ AML (FAB M0). The patient received induction chemotherapy as MA/VP plus Imatinib (400 mg/day×4w) and achieved CHR. Imatinib combined chemotherapy continued for another 7 more cycles. Imatinib was taken as 400 mg/day×2 w during the intermission period of chemotherapy. He had a bone marrow relapse with 18% blasts 48 days after the last chemotherapy, although CCyR and CMR were still maintained. Imatinib was added in a dose of 600mg/day and CHR achieved again 1 month later. The patient then took continuous Imatinib (400mg/day) until he received allo-SCT 26months after diagnosis. STR analysis showed completely donor phenotype 1, 3 and 6 months after SCT. Imatinib (400 mg/day) administration was continued and the patient maintained CHR and CMR in the following clinical course. However, he developed interstitial pneumonia 11 months after SCT and died from severe respiratory failure 30 months after SCT with CHR and CMR still maintained at that time. Our cases indicate that Imatinib combined with daunorubicin based chemotherapy followed by allo-SCT and Imatinib maintenance treatment appears to be the best way to treat Ph+ de novo AML, especially when Imatinib is used in an early phase of AML and when a complete molecular response is achieved before allo-SCT. Disclosures: No relevant conflicts of interest to declare.