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Background The nucleoporin 98 (NUP98)-paired related homeobox 1 (PMX1) fusion gene, which results from t(1;11)(q23;p15), is rare in patients with acute myeloid leukemia (AML). Currently, only two cases of chronic myeloid leukemia in the accelerated phase or blast crisis and three cases of therapy-related AML have been reported. Here, we first report a patient with de novo AML carrying the NUP98-PMX1 fusion gene. Case presentation A 49-year-old man diagnosed with AML presented the karyotype 46,XY,t(1;11)(q23;p15)[20] in bone marrow (BM) cells. Fluorescence in situ hybridization analysis using dual-color break-apart probes showed the typical signal pattern. Reverse transcription-polymerase chain reaction (RT-PCR) analysis suggested the presence of the NUP98-PMX1 fusion transcript. The patient received idarubicin and cytarabine as induction chemotherapy. After 3 weeks, the BM aspirate showed complete remission, and the RT-PCR result for the NUP98-PMX1 fusion gene was negative. Subsequently, the patient received three cycles of high-dose Ara-c as consolidation chemotherapy, after which he underwent partially matched (human leukocyte antigen–DP locus mismatch) unrelated allogeneic hematopoietic stem cell transplantation (HSCT). The follow-up period ended on September 30, 2020 (6 months after HSCT), and the patient exhibited no recurrence or transplantation-related complications. Conclusion This is the first report of a patient with de novo AML carrying the NUP98-PMX1 fusion gene. The reported case may contribute to a more comprehensive profile of the NUP98-PMX1 rearrangement, but mechanistic studies are warranted to fully understand the role of this fusion gene in leukemia pathogenesis.

BACKGROUND Pulmonary infection is one of the life-threatening complications occurring during allogeneic hematopoietic stem cell transplantation (alloHSCT), even when prophylactic measures have been employed. Few studies have investigated whether pulmonary infection affects platelet recovery during alloHSCT. MATERIAL AND METHODS We retrospectively reviewed 253 consecutive patients with hematologic diseases who received alloHSCT in our institute. Among them, 62 patients (25%) had pulmonary infection within 100 days after alloHSCT. Using the one-to-two propensity-score matching logistic model, 50 patients with pulmonary infection and 100 patients without were included based on age, disease and stage, time from diagnosis to transplantation, infused CD34⁺ cells, and mononuclear cells. RESULTS The incidences of prolonged thrombocytopenia in patients with pulmonary infection were 44% (22/50) and 9% (9/100) in the corresponding matched group (P

To investigate the effect of chronic graft-versus-host disease (cGVHD) on the outcomes of acute myeloid leukemia (AML) patients who relapsed after allogenic hematopoietic cell transplantation, we performed a retrospective analysis on 218 patients with a median follow-up of 21.4 (3.4–179.6) months. A total of 103 patients developed cGVHD, with a 2-year cumulative incidence of 48.9% (95% CI 42.1–55.7%). The estimated 3-year overall survival was 85.7% (95% CI 75.7–95.7%), 48.8% (95% CI 31.7–66.0%), and 54.1% (95% CI 44.3–63.8%) for patients with limited cGVHD, extensive cGVHD, and without cGVHD (P

Background: COVID-19 has taken a huge toll on medical resources and the economy and will inevitably have an impact on public mental health. Post-traumatic stress disorder (PTSD), as the most common mental illness after an epidemic, must be seriously addressed. This study aimed to investigate the subjective fear of the Chinese general public during COVID-19 and to explore how it affected the development of PTSD.Methods: An online questionnaire survey was conducted among 1,009 people from January 30 to February 14, 2020 (about 1 month after the COVID-19 outbreak). The subjective fear was measured by a self-reported single-choice question. Four items from the Pittsburgh Sleep Quality Index (PSQI) were selected to measure the subjects' sleep quality. Their post-traumatic stress symptoms (PTSS) were measured by the PTSD Checklist for DSM-5 (PCL-5). Pearson correlation, hierarchical multivariate regression analysis, multiple mediator model, and bootstrapping were used in statistical analyses.Results: Different people showed different levels of subjective fear in response to the outbreak. There was a significant positive correlation between subjective fear and the total score of PCL-5 (R = 0.513, P < 0.01), meaning that the higher the degree of subjective fear, the more severe the symptoms of post-traumatic stress are. Subjective fear was an important predictor of PTSS, accounting for 24.3% of the variance. The total effect of subjective fear on PCL-5 scores was significant (total effect = 7.426, SE = 0.405, 95% CI = 6.631–8.221). The total indirect effect of subjective fear on PCL-5 scores through sleep quality was also significant (total indirect effect = 1.945, SE = 0.258, 95% CI = 1.436–2.470).Conclusions: Subjective fear has an important predictive effect on PTSS. In addition to the direct effect, our findings firstly demonstrate the mediating role of sleep quality in the relationship between subjective fear and PTSS.

Introduction Relapse remains the leading cause of treatment failure after allogeneic hematopoietic stem cell transplantation (alloHSCT) in leukemia. Numerous investigations have demonstrated that minimal residual disease (MRD) before or after alloHSCT is prognostic of relapse risk. These MRD data were collected at specific checkpoints and could not dynamically predict the relapse risk after alloHSCT, which needs serial monitoring. Methods In the present study, we retrospectively analyzed MRD measured with multi-parameter flow cytometry in 207 acute myeloid leukemia (AML) patients (acute promyelocytic leukemia excluded), and 124 acute B lymphoblastic leukemia (ALL) patients. A three-step method based on joint model was used to build a relapse risk prediction model. Results The 3-year overall survival and relapse-free survival rates of the entire cohort were 67.1% ± 2.8% and 61.6% ± 2.8%, respectively. The model included disease status before alloHSCT, acute and chronic graft-versus-host disease, and serial MRD data. The time-dependent receiver operating characteristics was used to evaluate the ability of the model. It fitted well with actual incidence of relapse. The serial MRD data collected after alloHSCT had better discrimination capabilities for recurrence prediction with the area under the curve from 0.67 to 0.91 (AML: 0.66-0.89; ALL: 0.70-0.96). Conclusion The joint model was able to dynamically predict relapse-free probability after alloHSCT, which would be a useful tool to provide important information to guide decision-making in the clinic and facilitate the individualized therapy.

To compare the efficacy and toxicity of a novel regimen called FBA, consisting of fludarabine, busulfan, and cytarabine, with the standard BuCy2 regimen for younger adult patients with acute myeloid leukemia, we conducted a prospective randomized phase II study. Patients in complete remission were randomly assigned to receive either the FBA (n = 56) or the BuCy2 regimen (n = 55). The difference in 100-day transplant-related mortality (TRM) was not statistically significant between the two arms (1.79% for FBA versus 5.45% for BuCy2, P = 0.260), as were the cumulative incidences of relapse, TRM, overall survival (OS) and event-free survival (EFS) at 3 years. However, the 100-day cumulative incidences of grades II–IV and III–IV acute graft-versus-host disease (aGVHD) were lower in the FBA group [(8.93% versus 21.86%, P = 0.032) (1.79% versus 9.09%, P = 0.025)]. The 3-year GVHD and relapse-free survival (GRFS) was 31.20% for the FBA group and 14.96% for the BuCy2 group (P = 0.004). The incidences of diarrhea and severe oral mucositis within the first 30 days post-transplantation were lower in the FBA group [(28.57% versus 65.45%; P

Chronic graft-versus-host disease (cGVHD) is an autoimmune-like syndrome, and donor B cells play important roles in augmenting its pathogenesis. B cell–depleting anti-CD20 mAb has been administered before or after cGVHD onset for preventing or treating cGVHD in the clinic. Although administration before onset appeared to be more effective, the effect is variable and sometimes minimal. Here, we used 2 mouse cGVHD models to evaluate the preventive and therapeutic effect of anti-CD20 mAb. With the model of DBA/2 donor to MHC-matched BALB/c recipient, 1 intravenous injection of anti-CD20 mAb (40 mg/kg) the following day or on day 7 after hematopoietic cell transplantation when serum autoantibodies were undetectable effectively prevented induction of cGVHD and preserved a strong graft-versus-leukemia (GVL) effect. The separation of GVL effect from GVHD was associated with a significant reduction of donor CD4+ T cell proliferation and expansion and protection of host thymic medullary epithelial cells. Anti-CD20 mAb administration also prevented expansion of donor T cells and induction of cGVHD in another mouse model of C57BL/6 donor to MHC-mismatched BALB/c recipients. In contrast, administration of anti-CD20 mAb after GVHD onset was not able to effectively deplete donor B cells or ameliorate cGVHD in either model. These results indicate that administration of anti-CD20 mAb before signs of cGVHD can prevent induction of autoimmune-like cGVHD while preserving a GVL effect; there is little effect if administered after cGVHD onset. This provides new insights into clinical prevention and therapy of cGVHD with B cell–depleting reagents.

We retrospectively analyzed 449 patients with AML under the WHO classification of AML 2008 and probed implications of this classification in diagnosis and treatment of acute myeloid leukemia with myelodysplasia-related changes (AML-MRC) among them. The clinical presentations, biological features, treatments, and prognosis of patients diagnosed with AML-MRC were analyzed and compared with those of AML not otherwise specified (AML-NOS). In all patients, 115 (25.6%) were diagnosed as AML-MRC including 64 males and 51 females with median onset age of 48 years (range from 17 to 78). Their complete remission (CR) rate was 60.9% and relapse rate was 57.1%. The observed median overall survival (OS) and disease-free survival (DFS) were 10 and 5 months, respectively, which was significantly shorter than those of AML-NOS patients (P

Prevention of chronic graft-versus-host disease (cGVHD) remains a major challenge in allogeneic hematopoietic cell transplantation (HCT) owing to limited understanding of cGVHD pathogenesis and lack of appropriate animal models. In this study, we report that, in classical acute GVHD models with C57BL/6 donors and MHC-mismatched BALB/c recipients and with C3H.SW donors and MHC-matched C57BL/6 recipients, GVHD recipients surviving for >60 d after HCT developed cGVHD characterized by cutaneous fibrosis, tissue damage in the salivary gland, and the presence of serum autoantibodies. Donor CD8+ T cells were more potent than CD4+ T cells for inducing cGVHD. The recipient thymus and de novo–generated, donor-derived CD4+ T cells were required for induction of cGVHD by donor CD8+ T cells but not by donor CD4+ T cells. Donor CD8+ T cells preferentially damaged recipient medullary thymic epithelial cells and impaired negative selection, resulting in production of autoreactive CD4+ T cells that perpetuated damage to the thymus and augmented the development of cGVHD. Short-term anti-CD4 mAb treatment early after HCT enabled recovery from thymic damage and prevented cGVHD. These results demonstrate that donor CD8+ T cells cause cGVHD solely through thymic-dependent mechanisms, whereas CD4+ T cells can cause cGVHD through either thymic-dependent or independent mechanisms.

The prognosis of patients with refractory/relapsed acute myelogenous leukemia (rAML) is poor. Recent studies have shown that more transplant centers are choosing allogeneic peripheral blood stem cell transplantation (allo-PBSCT) for recipients, even with a higher leukemia burden. The purpose of the present study is to evaluate the outcome of rAML patients undergoing allo-PBSCT and to determine whether the disease status can predict the post-transplantation survival. The outcome of 58 patients (median age, 34 years; range, 14 to 52) with rAML who underwent allo-PBSCT in our institution from January 2000 until September 2011 was retrospectively studied. Thirty-three patients had complete remission (CR) before PBSCT, whereas 25 patients had no remission. Donors were matched related (31 patients) and unrelated (27 patients). Reduced-intensity conditioning was used for 18 patients with rAML, and myeloablative conditioning was used for others. Sixty-six consecutive non-rAML patients (median age, 33 years; range, 15 to 51) who received an allo-PBSCT at the same period were used as a control. Full donor-type engraftment was achieved in all patients. After a median follow-up of 61 months, the 5-year overall survival of rAML patients was 54.21% ± 7.06%, which was lower than non-rAML patients (71.82% ± 6.4%, P = .0386). However, the 5-year event-free survival for rAML and non-rAML patients had no statistical significance (53.54% ± 6.87% versus 62.07% ± 6.78%, P = .2626). The 5-year overall survival between rAML patients who had CR and no remission before PBSCT was 56.06% ± 9.2% and 51.85% ± 10.83%, respectively (P = .6408). These data demonstrate that allo-PBSCT is a promising and safe choice for the treatment of rAML, and the results were partially due to the rapid tapering of immunosuppressants in the early stage after PBSCT and prophylactic donor lymphocyte infusion. Meanwhile, the patients who did not achieve CR before PBSCT could also benefit from allo-PBSCT.

OBJECTIVE To evaluate the efficacy of rituximab combined with CHOP-like regimen with or without IFN in patients newly diagnosed diffuse large B-cell Non-Hodgkin’s lymphoma (DLBCL). METHODS From January 2003 to July 2008, 51 patients received CHOP-like chemotherapy (cyclophosphamide 750 mg/m2, epirubicin 80 mg/m2, vindesine 2.8 mg/m2 on day 1 , and prednisolone 100 mg/day on day 1 to day 5). Thirty- tne patients received CHOPR-like treatment (rituximab 375 mg/m2 1 day before CHOP-like chemotherapy). Twenty patients received CHOP-like regimen in combination with peginterferon (pegIFN) (1 μg/kg on day 5) and rituximab (on day 6). RESULTS The CR (complete remission) rate in the CHOPR-like (with or without pegIFN) group and in the CHOP-like group was 78.4% and 45.1% (P = 0.005), respectively. The estimated mean time of overall survival (OS) in the CHOPR-like group and CHOP-like group was 58.7 ± 2.8 and 36.4 ± 3.4 months, respectively (P = 0.002). The rates of CR and OR (overall remission) in CHOPR-like with IFN arm were 85.0% and 95.0%, and the rates of those in CHOPR-like without IFN arm were 74.2% and 87.0% (P > 0.05). The estimated mean time of 4-year-PFS (progression-free survival) in CHOPR-like with IFN arm and in CHOPR-like without IFN arm was 62.9 ± 3.0 months and 51.0 ± 4.6 months (P = 0.092), respectively. In the CHOPR-like with IFN arm, no patient relapsed after achieving CR, while the estimated rate of 4-year-DFS (disease-free survival) in the patients who reached CR in the CHOPR-like without IFN arm was (63.4 ± 19.3)% (P = 0.061). CONCLUSION Rituximab combined with CHOP-like chemotherapy improved the prognosis of DLBCL patients. The IFN may help to improve the quality and duration of response of DLBCL patients treated with rituximab and CHOP-like regimen.

The aim of the present study was to evaluate the effectiveness of bortezomib combined with epirubicin, dexamethasone, and thalidomide (BADT) for the treatment of multiple myeloma (MM). The BADT regimen consisted of a maximum of eight 4-week cycles of: intravenous bortezomib (1.0 mg/m(2)) and intravenous epirubicin (12 mg/m(2)) on days 1, 4, 8, and 11; dexamethasone (20 mg) on days 1, 2, 4, 5, 8, 9, 11, and 12; and oral thalidomide (100 mg/m(2)) on days 1-28. Twelve patients with MM were included in the study, of whom four had not been previously treated and eight had been previously treated with at least one cycle of a systemic combined regimen. All the patients completed at least two cycles of treatment, with an average of five cycles; the complete response (CR) rate was 83.3% (10/12) and stabilization of disease was 16.7% (2/12). The average number of cycles required to achieve CR was 1.9 (range 1-6). In three patients, mobilization of peripheral blood stem cells allowed a sufficient quantity of CD34+ cells to be harvested for future autotransplantation. The main adverse reactions included peripheral neuropathy (4/12), thrombocytopenia (3/12), electrocardiographic abnormalities (4/12), neutropenia (5/12), and liver function impairment (4/12), primarily grade I-II. Infection occurred in four patients with neutropenia, including one patient who developed sepsis. The estimated 1-year overall survival rate was 91.7 +/- 8.0%, and the estimated 1-year disease-free survival was 75.0 +/- 12.5%. BADT is a highly effective combined regimen, with acceptable toxicity, for the treatment of multiple myeloma.

Cutaneous sclerosis is one of the most common clinical manifestations of chronic graft-versus-host disease (cGVHD). Donor CD4(+) T and B cells play important roles in cGVHD pathogenesis, but the role of antibodies from donor B cells remains unclear. In the current studies, we generated immunoglobulin (Ig)H(µγ1) DBA/2 mice whose B cells have normal antigen-presentation and regulatory functions but cannot secrete antibodies. With a murine cGVHD model using DBA/2 donors and BALB/c recipients, we have shown that wild-type (WT) grafts induce persistent cGVHD with damage in the thymus, peripheral lymphoid organs, and skin, as well as cutaneous T helper 17 cell (Th17) infiltration. In contrast, IgH(µγ1) grafts induced only transient cGVHD with little damage in the thymus or peripheral lymph organs or with little cutaneous Th17 infiltration. Injections of IgG-containing sera from cGVHD recipients given WT grafts but not IgG-deficient sera from recipients given IgH(µγ1) grafts led to deposition of IgG in the thymus and skin, with resulting damage in the thymus and peripheral lymph organs, cutaneous Th17 infiltration, and perpetuation of cGVHD in recipients given IgH(µγ1) grafts. These results indicate that donor B-cell antibodies augment cutaneous cGVHD in part by damaging the thymus and increasing tissue infiltration of pathogenic Th17 cells.

Acute myeloid leukemia (AML) is generally regarded as a disorder of stem cells, known as leukemic initiating cells (LICs), which initiate the disease and contribute to relapses. Although the phenotype of these cells remains unclear in most patients, they are enriched within the CD34(+)CD38(-) population. In core-binding factor (CBF) AML, the cytogenetic abnormalities also exist in LIC. The aim of this study was to determine the prognostic power of minimal residual disease (MRD) measured by fluorescence in situ hybridization (FISH) in CD34(+)CD38(-) cells sorted by flow cytometry at different periods during therapy. Thirty-six patients under 65 years of age with de novo CBF-AML treated with intensive chemotherapy were retrospectively included in this study. Correlations with relapse-free survival (RFS) and overall survival were evaluated with univariate and multivariate analyses. FISH efficiently identified LICs in the CD34(+)CD38(-) population. The presence of FISH(+)CD34(+)CD38(-) cells before consolidation was negatively associated with cumulative incidence of relapse (64 vs 18 %, P = .012), which showed prognostic value for RFS (12 vs 68 %, P = .008) and OS (11 vs 75 %, P = .0005), and retained prognostic significance for RFS in multivariate analysis. The detection of FISH(+)CD34(+)CD38(-) cells before consolidation therapy significantly correlated with long-term survival. Fluorescence-activated cell sorting (FACS)-FISH could be potentially adopted as a MRD monitor approach in clinical practice to identify CBF-AML patients at risk of treatment failure during therapy.

Acute myeloid leukemia (AML) is a clonal hematologic malignancy arising from a small population of leukemic cells that initiate and propagate the disease. These cells are termed leukemic stem cells (LSCs), which are found within the CD34+CD38- cell compartment. The LICs subpopulation survives chemotherapy and is most probable the cause of minimal residual disease (MRD), which in turn is thought to cause relapse. The aim of this study was to determine the prognostic value of the percentage of LICs measured by fluorescence in situ hybridization (FISH) in flow sorted CD34+CD38- cells (FISH+CD34+CD38- population) at diagnosis. Forty-five patients with de novo AML with FISH-detectable cytogenetic abnormalities treated with CHAML 2010 protocol were retrospectively included in this study. The last follow-up was March 31, 2013, and the median follow-up for patients was 12 months (range: 1–27 months). After 24 months, 12 patients (26.7%) had experienced relapse, whereas 7 patients (17.9%) had died due to non-relapse treatment-related complications, 3 from treatment-related complications of transplantation and 4 from infection after chemotherapy. To distinguish normal and leukemic cells within the CD34+CD38- cell compartment, we established a unique protocol for conducting FISH on flow-sorted cells. The FISH positive percentage at diagnosis constituting an average of 2.31% (range: 0.01%-29.4%) of the blast cells and 68.2% (range: 7.8%-89.6%) of the CD34+CD38- cells. According to the LIC level detected by Flow-FISH analysis, patients were categorized into the two groups: low LIC load (＜1%) and high LIC load (≥1%), representing 44.4% (n=20) and 55.6% (n=25) of all patients, respectively. Comparison of clinical and laboratory characteristics showed no significant differences between two groups in age, gender, white blood cell (WBC) count, platelet (PLT) count, blast percentage, FAB subtype distribution, FLT3 mutation status and cytogenetics risk at diagnosis. High LIC load at diagnosis was significantly correlated with increased risk of poor clinical outcome. The 2-year overall survival (OS) for patients with low LIC load was 0.7257[0.4082, 0.8916], compared with 0.1675[0.0323, 0.3947] for the patients with high LIC load (P=0.0004). And 2-year events-free survival (EFS) were 0.6723[0.3262, 0.8687] versus 0.1633[0.0328, 0.3825], respectively (P=0.0029). By multivariate analysis, high LIC load retained prognostic significance for OS and EFS. Furthermore, high LIC load at diagnosis was found to be significantly associated with elevated cumulative incidence of relapse (2-year CIR: 56.7% vs. 18.0%; P = 0.021). In summary, our study established the Flow-FISH protocol as a useful method to distinguish normal and leukemic cells within the CD34+CD38- cell subpopulation. And Flow-FISH might be easily adopted as a powerful tool to predict clinical outcome and help physicians to evaluate criteria for treatment in AML with recurrent cytogenetic abnormalities. Disclosures: No relevant conflicts of interest to declare.

Recipient tissue expression of programmed death-ligand 1 (PD-L1, B7H1) down-regulates graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation (HCT), but this information has not been translated clinically. Here we demonstrate a critical role for recipient PD-L1 expression in preventing both acute and chronic GVHD while preserving graft-versus-leukemia (GVL) effects when donor CD4+ T cells are temporarily depleted in vivo early after HCT. Depletion of donor CD4+ T cells increases serum IFN-γ concentrations and enhances recipient tissue expression of PD-L1 and donor CD8+ T cell expression of the PD-L1 receptors CD80 and PD-1. In GVHD target tissues, depletion of CD4+ T cells increases anergy and apoptosis of infiltrating CD8+ T cells in a manner that depends on recipient PD-L1 expression, thereby preventing damage to intestinal Paneth cells and stem cells, hepatocytes, and thymic medullary epithelial cells, and preventing both acute and chronic GVHD. In lymphoid tissues, depletion of CD4+ T cells augments CD8+ T cell proliferation without increasing CD8+ T cell anergy or apoptosis, resulting in expansion of donor CD8+ T cells and strong GVL effects. Therefore, temporary depletion of donor CD4+ T cells early after HCT represents a novel approach for preventing GVHD while preserving GVL effects. This work was supported by NIH R01 AI066008 and Nesvig Lymphoma Fundation (to D.Z.) and by NSFC 81090413, 81270638 (to J.W.). Disclosures Forman: Amgen: Consultancy; Mustang: Research Funding. Martin:Neovii: Research Funding; RegImmune: Research Funding; Enlivex: Consultancy; Janssen: Consultancy; Pfizer: Consultancy; Pharmacyclics: Consultancy.

Background Biphenotypic acute leukemia is a rare disorder that is difficult to diagnose. It displays features of both myeloid and lymphoid lineage. There is still a lack of studies in biphenotypic acute leukemia in a Chinese population. We present here a comprehensive investigation of the clinical and biological characteristics, and outcome of biphenotypic acute leukemia in our hospital in over a seven year period. Design and Methods We retrospectively analyzed 452 adult acute leukemia patients diagnosed according to French-American-British (FAB) classification and biphenotypic acute leukemia diagnosed according to European Group for the Immunological Characterization of Leukemias (EGIL) classification, respectively. Biological characteristics, response to treatment, and outcome were examined in biphenotypic acute leukemia patients and compared with that in acute myeloid leukemia and acute lymphoblastic leukemia patients with complete follow-up profiles diagnosed in the same period. Results Of 452 acute leukemia patients, 21 cases (4.6%) were diagnosed as biphenotypic acute leukemia. Among them, 14 (66.7%) were B lymphoid and myeloid, 5 (23.8%) were T lymphoid and myeloid, one (4.8%) was T/B lymphoid and one (4.8%) was trilineage differentiation. When compared with acute myeloid leukemia and acute lymphoblastic leukemia, patients with biphenotypic acute leukemia showed significantly higher incidence of CD34 antigen expression, unfavorable karyotypes, and extramedullary infiltration ( p

We recently reported that in a chronic graft versus host disease (GVHD) model of DBA/2 donor to MHC-matched BALB/c recipient, donor CD4+ T and B cell interaction resulted in not only hyperglobulinemia and glomerulonephritis but also scleroderma (J. Immunol. 2012). It is well known that glomerulonephritis is caused by immune complex deposition. However, the role of antibodies from donor B cells in the pathogenesis of scleroderma remains unclear. To address this question, we generated DBA/2 mice whose B cells have APC function but cannot secrete antibodies by backcrossing IgHµg1 mice from Dr. Rajewsky’s lab (JEM 2007). We observed that, while transplanting T-cell-depleted bone marrow (TCD-BM) and spleen cells from littermate control mice induced proteinuria and scleroderma, transplanting BM and spleen cells from IgHµg1 DBA/2 mice induced no proteinuria, but the recipients developed scleroderma ~35 days after HCT. Interestingly, the scleroderma gradually recovered ~55 days after HCT. 40 days after HCT, scleroderma recipients transplanted with WT spleen cells (Rec-WT) or recipients transplanted with IgHµg1 spleen cells (Rec-IgHµg1) both had high percentage (~12%) of IFN-g+ or IL-17+ CD4+ T cells in the peripheral lymph node (PLN) and skin tissues, as compared to that (~3%) of GVHD-free recipients given TCD-BM alone (Rec-TCD). While Rec-WT had severe reduction of CD4+CD8+ thymocytes, the Rec-IgHµg1 had no reduction of the thymocytes, as compared to that of Rec-TCD. By day 60 after HCT, the Rec-WT with ongoing scleroderma still had ~10% IFN-g+ or IL-17+ CD4+ T cells in the PLN and skin tissues; in contrast, although the Rec-IgHµg1 with reversal of scleroderma still had >10% IFN-g+or IL-17+ CD4+ T cells in the PLN, those cells in the skin had reduced to Disclosures No relevant conflicts of interest to declare.

Donor T cell-mediated graft versus leukemia/lymphoma (GVL) effect plays a critical role in preventing tumor relapse in leukemia/lymphoma patients treated with allogeneic hematopoietic cell transplantation (HCT). However, these same donor T cells also induce acute and chronic graft versus host disease (GVHD), which remains a major obstacle for allogeneic HCT as a curative therapy for hematological malignancies. Chronic GVHD is a systemic lupus- and scleroderma-like autoimmune syndrome. We and others reported that donor B cells play important roles in augmenting pathogenesis of chronic GVHD in mouse models and humans. Rituxan (an anti-CD20 mAb) has been used in the clinic for the treatment of ongoing chronic GVHD, but the effect is variable and minimum in some reports, and the mechanisms for this ineffectiveness remains unclear. In the current studies, we evaluated the effect of in vivo administration of a depleting anti-CD20 mAb in preventing and treating autoimmune-like chronic GVHD as well as the impact on GVL effect. With the mouse chronic GVHD model of DBA/2 donor to MHC-matched BALB/c recipient established in our lab (Blood, 2006, J. Immunol 2012), we found that one intravenous injection of anti-CD20 (50mg/kg) immediately after HCT effectively prevented induction of autoimmune-like chronic GVHD. While all (12/12) recipients treated with control IgG developed chronic GVHD with proteinuria and hair-loss and died by 30 days, none (0/12) of the anti-CD20-treated recipients developed proteinuria or hair-loss and all survived for more than 50 days after HCT without signs of chronic GVHD. In addition, the anti-CD20-treated recipients eliminated BCL1 leukemia/lymphoma cells without signs of chronic GVHD. The preventative anti-CD20 treatment had little impact on donor CD8+ T cell activation and expansion in the periphery and allowed for the strong CD8+ T cell-mediated GVL effect. The effective prevention of chronic GVHD by anti-CD20 was associated with significant changes in donor B and CD4+ T cells as well as associated with protection of host thymus. Preventive anti-CD20 treatment depleted IL-6-producing donor B cells and increased IL-10-producing B cells. In addition, the treatment also significantly reduced donor CD4+ T cell expansion, the percentage of IFN-g-producing CD4+ T as well as CD4+ T cells expressing CD8αα, the latter of which has recently been reported to represent over-activated pathogenic CD4+ T cells, such that the host thymus was protected from donor T-cell-mediated damage. Anti-CD20 or control IgG treatment of recipients with ongoing chronic GVHD 15-20 days after HCT did not show any difference in disease progress, and all (8/8) in each group died within 30 days after HCT. The ineffectiveness of anti-CD20 therapy was associated with little reduction of CD19+ B or CD4+ T cells, as the majority of CD19+ B cells became CD20- in the recipients with ongoing chronic GVHD. These results indication that anti-CD20 can effectively prevent induction of autoimmune-like chronic GVHD while preserving the GVL effect, but anti-CD20 is ineffective in treating ongoing chronic GVHD. (This study is supported by the Nesvig Lymphoma Foundation). Disclosures: Chan: Genentech Inc.: Employment.

Promising efficacy of bortezomib, the first therapeutic proteasome inhibitor, has been demonstrated in the treatment of multiple myeloma (MM). In this study, we evaluate the therapeutic effects and adverse reactions of bortezomib combination with epirubicin, dexamethasone and thalidomide (BADT) for the treatment of multiple myeloma (MM). Using BADT regimen (bortezomib 1.0mg/m2 on days 1, 4, 8 and 11; epirubicin 12mg/m2 on days 1, 4, 8, 11; dexamethasone 20mg on days 1–2, 4–5, 8–9, 11–12; thalidomide 100mg/m2 on days 1–28) for a maximum of eight 4-week cycles. 12 MM patients were treated, including 5 previously untreated patients and 7 previously treated patients. Each patient completed at least 2 cycles with a mean of 4.5 cycles. Of the 12 patients, 9 patients (75%) achieved complete remission (CR)and 2 patient (16.7%) achieved partial remission (PR), with an overall response (OR) of 91.5%. The mean number of cycles to CR was 2.8 (1–6), and the mean number of cycles to PR was 1.3 (1–3). 7 patients achieved PR after one cycle treatment, and 4 patients achieved CR after one cycle treatment. The main adverse reactions included peripheral neuropathy (4/12), thrombocytopenia (3/12), abnormal change of ECG (4/12), neutropenia (5/12), and liver function impairment (3/12), mainly grade ±-II. Infection occurred in three patients with neutropenia, including sepsis in one patient and herpes zoster in another patient. One patient died of diseases progression, another patient died of secondly sarcoma after achievement CR. The median progression-free time was 14 months (2–26) months, including 2 patients who have been in CR for 15 months since discontinuation of the treatment. In Conclusion, the BADT regimen is highly effective with low toxicity in the treatment of MM. It is warranted to conduct further study on this regimen in more cases.

Objective To explore the significance of reduced-intensity conditioning allogeneic peripheral blood hematopoietic stem cell transplantation (PBSCT) in the treatment of chronic myeloid leukemia. Patients and Methods Twenty-six consecutive patients received reduced-intensity conditioning (Flu 30mg/m2.d−1×5d+BU 4mg.kg−1.d−1×3d or CTX 100mg.kg−1+TBI≤6.0Gy) PBSCT(RIC group). Among them, 21 were male and 5 female with median age of 36 (23~49). Twenty three of them were in chronic phase and 3 in progressive phases. The median time from diagnosis to transplantation was 12 (3~84) months. Twenty-four consecutive patients received standard conditioning (CTX 120mg.kg−1+ TBI≥7.0Gy) prior to PBSCT (STAND group) were used as historic controls. In this group of patients, 22 were in chronic phase and 2 in progressive phases. Twenty patiens were male and 4 female with median age of 35 (18~49). The median time from diagnosis to transplantation was 13 (3~48) months. All patients received PBSC from HLA matched related donors. Mycophenolate mofetil (MMF),CsA and MTX were given for prophylaxis of acute graft-versus-host-disease (aGVHD). Results All patients were successfully engrafted and achieved a complete cytogenetic remission. One patient developed graft rejection 6 months post-transplantation in RIC group. The median time when granulocyte exceeded 0.5 ×109/L and platelets exceeded 20 ×109/L was 15(12–23) days and 19(12–32) days in STAND group and 13(11–18) and 17(11–30) days in RIC group, respectively. The cumulative incidence of aGVHD was 45.83% (11/24) in STAND group and 23.08% (6/26) in RIC group(P>0.05). The incidence of Grade III–IV aGVHD in STAND group and RIC group was 16.67% and 0%, respectively(P0.05). The incidence of extensive cGVHD in STAND group and RIC group was 35% and 0%, respectively(P0.05). 4 patients experienced a cytogenetic relapse, which was successfully treated with donor PBSC infusions. The Cumulative incidence of TRM was 41.67% (10/24) for the STAND group and 19.23% (5/26) for the RIC group, respectively (P>0.05). GVHD copplicated with interstitial pneumatitis or severe infection were the main causes of death. The estimated 3-year probabilities of disease-free-survival (DFS) was 62% in STAND group and 76% in RIC group, respectively (P>0.05). Conclusion Our results indicate that reduced-intensity conditioning allogeneic peripheral blood stem cells transplantation is a safe, less toxic and curative approach for patients with chronic myeloid leukemia.