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We aimed to evaluate the efficacy and safety of allogeneic hematopoietic stem cell transplantation with targeted oral busulfan (BU) and cyclophosphamide (CY) in a phase II study. Busulfan (1.0 mg/kg) was given initially in six doses. Based on the estimated concentration at steady state after the first dose of BU, subsequent (7th-16th) doses were adjusted to obtain a targeted overall concentration at steady state of 700-900 ng/mL. The primary endpoint was 1-year overall survival (OS). Fifty patients were registered and 46 (median age, 53 years; range, 18-62 years) received planned transplant, including 24 with AML, 16 with myelodysplastic syndrome, and six with CML. Fourteen patients were categorized as standard risk. Nineteen patients received transplant from human leukocyte antigen-identical siblings, 27 from unrelated donors. The BU dose required reduction in 32 patients and escalation in six patients. One-year OS was 65% (95% confidence interval, 50-77%). Cumulative incidence of hepatic sinusoidal obstruction syndrome was 11%. One-year transplant-related mortality was 18%. Both OS and transplant-related mortality were favorable in this study, including patients of older age and with high risk diseases. Individual dose adjustment based on BU pharmacokinetics was feasible and effective in the current phase II study. This trial is registered in the University Hospital Medical Information Network Clinical Trial Registry System (UMIN-CTR, ID:C000000156).

Adult T-cell leukemia/lymphoma (ATLL) is a highly aggressive hematologic neoplasm that has an extremely poor prognosis; however, this has improved following recent progress in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Several clinical studies have shown that discontinuation of immunosuppressant therapy induces durable remission in a significant number of post-transplant relapsed patients, suggesting that ATLL may be susceptible to a graft-versus-leukemia effect. Here, we report two cases with ATLL who received donor lymphocyte infusions (DLIs) for relapse after allo-HSCT; one patient achieved complete remission (CR) after a single DLI, and the other suffered repeated relapses and was treated with chemotherapy and radiotherapy combined with a total of five rounds of DLIs. Both patients presented with exacerbation of the graft-versus-host disease after the DLIs, and remained in CR for 9 and 8 years, respectively. These data support the use of DLIs as an effective therapy to induce durable CR in the treatment of relapsed ATLL. In this study, we review previous reports and discuss the role of DLIs in the treatment of post-transplant relapsed ATLL.

Allogeneic hematopoietic cell transplantation (HCT) is an effective treatment for adult T-cell leukemia (ATL), raising the question about the role of graft-versus-leukemia effect against ATL. In this study, we retrospectively analyzed the effects of acute and chronic graft-versus-host disease (GVHD) on overall survival, disease-associated mortality, and treatment-related mortality among 294 ATL patients who received allogeneic HCT and survived at least 30 days posttransplant with sustained engraftment. Multivariate analyses treating the occurrence of GVHD as a time-varying covariate demonstrated that the development of grade 1-2 acute GVHD was significantly associated with higher overall survival (hazard ratio [HR] for death, 0.65; P = .018) compared with the absence of acute GVHD. Occurrence of either grade 1-2 or grade 3-4 acute GVHD was associated with lower disease-associated mortality compared with the absence of acute GVHD, whereas grade 3-4 acute GVHD was associated with a higher risk for treatment-related mortality (HR, 3.50; P < .001). The development of extensive chronic GVHD was associated with higher treatment-related mortality (HR, 2.75; P = .006) compared with the absence of chronic GVHD. Collectively, these results indicate that the development of mild-to-moderate acute GVHD confers a lower risk of disease progression and a beneficial influence on survival of allografted patients with ATL.

Virus-associated hemorrhagic cystitis (HC) is a major cause of morbidity and mortality following allogeneic hematopoietic stem cell transplantation (HSCT). Although numerous studies have attempted to identify factors that predispose patients to viral HC, its causes remain controversial. We analyzed retrospectively the results of 266 allogeneic HSCTs to identify factors associated with HC. Of this group, 42 patients (15.8%) were diagnosed with viral HC, because of either adenovirus (ADV; n = 26; 9.8%) or BK virus (BKV; n = 16; 6.0%). ADV-HC was frequently associated with T cell purging, and was less common in patients with acute graft-versus-host-disease (GVHD). Conversely, BKV-HC was more frequently observed in patients with excessive immune reactions such as GVHD, preengraftment immune reaction, and hemophagocytic syndrome. These observations indicate that ADV- and BKV-HC may differ significantly in their risk factors and pathogenesis. Profound immune deficiency is more likely to be associated with ADV-HC, whereas immune hyperactivity might play a key role in BKV-HC.

We present a case of amebic colitis and liver abscess complicated by acute myeloid leukemia (AML) with high serum procalcitonin (PCT). A 61-year-old Japanese man seen at our hospital for severe diarrhea and high fever was found to have multiple ulcers in the transverse and sigmoid colon and rectum by colonoscopy and biopsies were conducted. Immature leukocytes with mild anemia and thrombocytopenia were seen in peripheral blood, necessitating bone marrow aspiration and biopsy that yielded a diagnosis of AML (FAB M4Eo). Serum C-reactive protein and PCT were extremely elevated. Blood cultures for bacteria and fungi were negative. Multiple low-density areas in the liver were found in abdominal computed tomography. Histological colon biopsy findings revealed amebic colitis, strongly suggesting amebic liver abscess. Metronidazole treatment was initiated for amebiasis and subsequent standard chemotherapy for AML was followed after fever was lowered. Hematological and cytogenetic CR was maintained with good clinical condition. Few case reports have been published in Japan to date on amebic colitis and liver abscess complicated by AML and no reports have been made on PCT elevation caused by amebiasis. In conclusion, differential diagnosis of amebiasis is necessary in addition to that of bacterial or fungal infection in serum PCT elevation.

The aim of this study is to evaluate the mechanism of long-term effect of autologous haematopoietic stem cell transplantation (ASCT) in treatment of SSc.Eleven patients (three males and eight females) with SSc were enrolled. Blood mononuclear cells were harvested after mobilization treatment with CYC and G-CSF. CD34+ haematopoietic stem/progenitor cell fractions were purified and cryopreserved. Patients were transplanted with2 × 10(6)/kg autologous CD34+ cells after high-dose CYC (50 mg/kg for 4 days) conditioning. Immune reconstitution was evaluated serially by analysing lymphocyte subpopulations for 36 months.Progressive improvement of skin sclerosis has been observed for 3 years in most of the patients. The serum level of anti-Scl-70, an auto-antibody specific to SSc, was progressively decreased after ASCT. Improvement of skin sclerosis was significantly associated with the change in the serum anti-Scl-70 level after ASCT at 36 months. Serum levels of KL-6 and surfactant protein D, indicators for interstitial pneumonia activity, were also significantly decreased. The number of CD8+ T cells immediately recovered within a month after ASCT, while the number of CD4+ T cells remained low for36 months post-transplant. The majority of CD4+ cells were memory but not naïve T cells, and regulatory CD4+ T cells were not recovered. Th1/Th2 ratio was significantly increased after ASCT.ASCT with purified CD34+ cells was effective in controlling the disease activity of SSc. Th1/Th2 ratio was significantly increased for at least 3 years after ASCT.

Introduction A clinical indication for allogeneic hematopoietic stem cell transplantation (HSCT) in adult Philadelphia-chromosome negative [Ph (-)] acute lymphoblastic leukemia (ALL) patients in complete remission 1 (CR1) remains to be clarified. Minimal residual disease (MRD) status has been reported to be a strong prognostic factor in adult ALL patients (Blood. 2006 107:1116, 2009 113:4153)( J Hematol Oncol. 2013 6:14). We prospectively monitored MRD status during induction and consolidation therapy in adult Ph (-) ALL patients to determine a clinical indication for HSCT. Materials & Methods From December 2008 to November 2013, 103 adult ALL patients were enrolled in this study. Eligibility criteria included non-L3 ALL, 16-65 years of age, and adequate organ function. Of these patients, 95 were eligible for this study and 59 were Ph (-). The treatment protocol used in this study was modified CALGB 19802. Treatment consisted of 6 courses given in the order of A-B-C-A-B-C, followed by a maintenance phase. Induction chemotherapy (course A) consisted of cyclophosphamide (1,200 mg/m2), daunorubicin (60 mg/m2 x 3), vincristine (VCR) (1.3 mg/m2 (max 2mg) x 4), L-asparaginase (3,000 U/m2 x 6) and prednisolone. Granulocyte colony-stimulating factor was started on day 4 and continued until neutrophil recovery. Consolidation B consisted of mitoxantrone (10 mg/m2 x 2), cytarabine (2,000 mg/m2/day x 4) and intrathecal (IT) administration of methotrexate (MTX). Consolidation C consisted of VCR (1.3 mg/m2 (max 2mg) x 3) and MTX (1,500 mg/m2 x 3) with leucovorin rescue and IT MTX. Patients received maintenance chemotherapy on a monthly basis: prednisolone 60 mg/m2 on days 1-5, VCR (1.3mg/m2 (max 2mg) x 3) on day 1, oral MTX 20 mg/m2 weekly, and oral 6-mercaptopurine 60 mg/m2 daily. MRD status was evaluated after induction therapy (first course A) and after second consolidation therapy (first course C). When MRD statuses after the consolidation were positive, patients were supposed to proceed to HSCT whenever possible. Results Among the 59 Ph-negative ALL patients, 51 patients (86%) could be monitored for MRD status, and the remaining 8 patients were not because of no clonal TCR/Ig targets or chimeric mRNA. The MRD status was determined by PCR analysis of major gene rearrangements and/or chimeric mRNAs (18 were positive for TCRγ, 16 for TCRδ, 13 for IgH, 1 for TCRγ and MLL-AF4, 1 for TCRγ and ETV6-RUNX1, 1 for IgH and MLL-AF4, and 1 for IgH and MLL-ENL). MRD quantifications were performed using ASO-primers with a sensitivity of ≤1 × 10−4, and MRD positivity was defined as a lower limit of detection of ≥1 × 10−3. The median follow-up time was 1597 days (range, 16-2870 days). A total of 51 patients included 29 males and 22 females, and their median age was 29 years ranging from 16 to 65. The median white blood cell count at presentation was 8.5 × 103/L (range 1.2-650.4). CR was achieved in 45 patients (88%). One patient died during induction due to intestinal bleeding. There were 29 survivors after the median follow-up period. The probability of 3-year OS and DFS in these patients with Ph-negative ALL was 69% (95%CI 54-80) and 50% (95%CI 36-63, respectively. In terms of CR1 status, MRD-negative patients after induction chemotherapy A in the first course (n = 17) showed a better 3-year DFS (65%) compared with MRD-positive patients (n = 31; 43%), as shown in Figure 1. The difference was not statistically significant (p = 0.07). There was no patient who proceeded to allogeneic HSCT among 17 MRD-negative patients after induction therapy in CR. In contrast, patients who were MRD-positive after induction but became MRD-negative after consolidation chemotherapy C in the first course (n = 11) showed a worse 3-year DFS compared with patients who were MRD-negative after induction chemotherapy A in the first course (45% vs. 65%, p = 0.08). Fourteen patients were MRD-positive after consolidation chemotherapy C in the first course. Among them, 5 patients proceeded to allogeneic HSCT in 1CR, and 9 did not. Three-year DFS with or without allogeneic HSCT were 60% (95%CI 13-88) vs 44% (95%CI 14-72, p=0.52), respectively. Discussion The present study indicates that MRD status after induction and consolidation therapies is a strong prognostic factor. Post-induction MRD-negative patients have been identified to have good-prognosis with chemotherapies, not suitable for up-front allogeneic HSCT. Figure 1. Figure 1. Disclosures Takamatsu: Taisho Toyama Pharmaceutical: Research Funding; TAIHO Pharmaceutical: Research Funding; Pfizer: Research Funding; Bristol-Myers Squibb: Research Funding; Ono Pharmaceutical: Research Funding; Astellas Pharma: Research Funding; Kyowa Hakko Kirin: Research Funding; Chugai Pharma: Research Funding; Takeda Pharmaceutical: Research Funding; Celgene: Honoraria. Akashi:Celgene: Research Funding, Speakers Bureau; Novartis pharma: Research Funding; Ono Pharmaceutical: Research Funding; Eisai: Research Funding; sanofi: Research Funding; Pfizer: Research Funding; Chugai Pharma: Research Funding; Kyowa Hakko Kirin: Research Funding, Speakers Bureau; Asahi-kasei: Research Funding; MSD: Research Funding; Otsuka Pharmaceutical: Research Funding; Taiho Pharmaceutical: Research Funding; Bristol-Myers Squibb: Research Funding, Speakers Bureau; Astellas Pharma: Research Funding; Eli Lilly Japan: Research Funding.

[Introduction] Allogeneic hematopoietic stem cell transplantation (Allo-SCT) remains the only potentially curative therapy for patients with high-risk or chemo-refractory hematologic malignancies. Recently, various methods including post-transplant cyclophosphamide (PT-CY) showed the ability to overcome the HLA disparity barrier and haploidentical hematopoietic stem cell transplantation (haplo-SCT) have been becoming an attractive method of transplantation with less rejection or graft-versus-host disease (GVHD). There have been many reports that the outcomes of haplo-SCT are equivalent to those of matched related donors and matched unrelated donors. The standard dose of PT-CY is 50 mg/kg/day for 2 days. However, optimal dose of PT-CY has not been studied extensively. In this study, we performed a clinical study of reduced dose of PT-CY (40 mg/kg/day for 2 days) haplo-SCT for hematologic malignancies. [Methods] We included adult patients with hematologic malignancies who received haplo-SCT at Kurume university hospital, Kurume, Japan between Oct 2014 and March 2018. Myeloablative conditioning (MAC) regimen consisted of fludarabine (Flu) (30 mg/m2 for 5 days), busulfan (BU) (3.2 mg/kg/day for 4 days) and total-body irradiation (TBI) 4Gy (Flu/BU4/TBI4Gy) or Flu (30 mg/m2 for 3 days) and TBI 12Gy (Flu/TBI12Gy). Reduced-intensity conditioning (RIC) regimen consisted of Flu (30 mg/m2 for 5 days), BU (3.2 mg/kg/day for 2 days) and TBI 4Gy (Flu/BU2/TBI4Gy). All patients were given PT-CY (40 mg/kg/day) on day +3 and day +4, followed by tacrolimus and mycophenolate mofetil (MMF) starting on day + 5 for GVHD prophylaxis. If there was no active GVHD, MMF was tapered off from day +30. Filgrastim 300 ug/m2 was administered starting on day +5 and continuing until neutrophil engraftment was achieved. Donors were mobilized with filgrastim 400 ug/m2 for 4 or 5 days, the peripheral blood stem cells were collected with one or two apheresis procedures. OS and PFS were calculated using the Kaplan-Meier method and the log-rank test was used for comparisons of Kaplan-Meier curves. Estimates of acute GVHD was calculated using death as the competing risk. P-values [Results] A total 15 patients with acute myeloid leukemia (AML, n=6), myelodysplastic syndrome (MDS, n=2), acute lymphoblastic leukemia (ALL, n=4), adult T-cell leukemia/lymphoma (ATLL, n=1) and malignant lymphoma (ML, n=2) were analyzed in this study. Patients median age was 54 years (range, 17 to 72); 9 patients were male, and 6 patients were female. Eight patients received MAC, 7 received RIC conditioning regimen. Disease status at transplantation was complete remission (CR) in 8 patients, non-CR in 7. The median CD34+ cell dose was 5.9 x 106 /kg (range, 2.4 to 17.4). All 15 patients achieved a neutrophil engraftment in a median 15 days (range, 13 to 19). Grade II-IV and III-IV acute GVHD occurred in 46.7% (95% confidence interval [CI], 14.4-66.8) and in 13.3% (95% CI, 0-28.9) patients, respectively. At 2 years, overall survival (OS) and progression-free survival were 36.6% (95% CI, 13.0-60.9) and 32.0% (95% CI, 10.9-55.7), respectively. The 2-years OS of patients in CR before SCT was 50.0% (95% CI, 15.2-77.5) compared to 28.6% (95% CI, 4.1-61.2) not in CR (P=0.106). The causes of death were: disease progression (n = 3), infection (n = 3), acute GVHD (n = 2), and noninfectious pulmonary complications (n = 2). Among infectious complications, two were bacterial infections and one was BK virus-associated hemorrhagic cystitis. [Conclusion] Reduced dose of PT-CY HLA haploidentical peripheral blood stem cell transplantation resulted in acceptable rate and severity of acute GVHD. However, relapse and infection remain major problems after PT-CY haplo-SCT for hematologic malignancies. Disclosures No relevant conflicts of interest to declare.

Background: High incidence of graft failure is a major obstacle for successful unrelated cord blood transplantation (UCBT). To ensure engraftment, unrelated cord blood (UCB) unit with higher total nucleated cell (TNC) and CD34 cell doses and less HLA mismatches between UCB units and patients were usually selected. However, it remains unclear whether HLA haplotype between the UCB units and patients should be matched. Using data from Japanese registry, we analyzed the effect of haplotype matching on the outcomes after single UCBT. Methods: Patients with hematologic diseases, aged 16 years or more, and have undergone their first UCBT between 2005 and 2014 were included in the study. The effect of haplotype matching (0, 1, and 2 haplotype matches) on neutrophil engraftment, overall mortality, relapse, and non-relapse mortality was analyzed after adjusting for other significant variables. Haplotype was estimated based on HLA-A, -B, -C, and -DRB1 allele information and Japanese allele frequency data. The haplotypes of both the patient and UCB unit were determined in 1,347 cases. Cases with more than 4 allele mismatches (n = 96) were excluded from the study. Results: The median age of the patients included in the study was 55 years (range 16-79 years). Median TNC and CD34 doses were 2.7 x 107/kg (range, 1.4-8.1x107/kg) and 0.8 x 105/kg (range, 0.1-5.2x105/kg), respectively. The number of allele mismatches were 0 in 82, 1 in 154, 2 in 252, 3 in 565, and 4 in 294 patients. The graft-versus-host disease (GVHD) prophylaxes were calcineurin inhibitors in combination with methotrexate in 707, calcineurin inhibitors in combination with Mycophenolate mofetil in 430, and other prophylaxes in 208 patients. The cumulative incidence of neutrophil engraftment at day 42 among groups with 0, 1, and 2 HLA haplotype matches was 79%, 82%, and 88%, respectively (Gray test, P=0.008) (Figure 1). In the multivariate analysis, the group with no haplotype match was marginally associated with worse neutrophil engraftment compared to the group with 1 haplotype match group (0 match vs. 1 match, HR 0.88, P=0.087), whereas the group with 2 haplotype matches was significantly associated with better neutrophil engraftment (2 match vs. 1 match, HR 1.39, P=0.005). Other significant variables were recipient sex, CD34 cell dose, transplant year, performance status, ABO matching, GVHD prophylaxis, and disease status. Tendency of worse neutrophil engraftment in the group with no haplotype matches was more apparent in patients with double mismatches at the same locus (0 match + no double mismatch vs. 1 match, HR 0.93, P=0.429, 0 match + double mismatch vs. 1 match, HR 0.80, P=0.050). Haplotype matching did not have any effect on overall survival and non-relapse mortality. Group with 2 haplotype matches was significantly associated with higher risk of relapse than the group with 1 haplotype match, whereas, there was no difference in the risk of relapse between the groups with no match and 1 match. Conclusions: In addition to the CD34 cell dose and HLA matching, HLA haplotype matching might be considered to obtain better neutrophil engraftment. Two haplotype matches (8 of 8 allele matches) should be avoided if the relapse risk is high. Figure 1. Figure 1. Disclosures Tanaka: Pfizer: Honoraria; Otsuka: Honoraria; Bristol-Myers Squibb: Honoraria, Research Funding; Novartis Pharma: Honoraria. Ichinohe:Zenyaku Kogyo Co.: Research Funding; Celgene: Honoraria; Janssen Pharmaceutical K.K.: Honoraria; Taiho Pharmaceutical Co.: Research Funding; Mundipharma: Honoraria; Otsuka Pharmaceutical Co.: Research Funding; Nippon Shinyaku Co.: Research Funding; Sumitomo Dainippon Pharma Co.: Research Funding; MSD: Research Funding; Novartis.: Honoraria; JCR Pharmaceuticals: Honoraria; Alexion Pharmaceuticals: Honoraria; Repertoire Genesis Inc.: Research Funding; Bristol-Myers Squibb: Honoraria; Takeda Pharmaceutical Co.: Research Funding; Pfizer: Research Funding; Ono Pharmaceutical Co.: Research Funding; Kyowa Hakko Kirin Co.: Research Funding; Eisai Co.: Research Funding; CSL Behring: Research Funding; Chugai Pharmaceutical Co.: Research Funding; Astellas Pharma: Research Funding.

Bortezomib and doxorubicin have synergistic activity against myeloma cells in vitro. We underwent a dose finding study of bortezomib in combination with a fixed dose of doxorubicin and intermediate-dose dexamethasone (iPAD therapy) in patients with relapsed or refractory myeloma. Bortezomib was administered on days 1, 4, 8 and 11 at a dose of 1.0 and 1.3 mg/m² in cohorts 1 and 2, respectively. Doxorubicin 9 mg/m² was given by rapid intravenous infusion on days 1-4, and dexamethasone 20 mg on days 1-2, 4-5, 8-9 and 11-12. Treatment was repeated at a 3-week interval and the dose-limiting toxicity (DLT), defined as grade 4 hematological toxicity lasting more than 5 days and/or grade 3 or higher non-hematological toxicity, was evaluated. In cohort 1, 2 of 6 patients developed DLTs including grade 4 hyponatremia and grade 3 infection with appropriate neutrophil counts. No DLT was observed in the remaining 4 patients, indicating this dose was tolerable. In cohort 2, 3 of 5 patients developed DLTs including grade 4 thrombocytopenia lasting more than 5 days, grade 3 hepatic transaminase elevation and grade 3 ileus, indicating this dose was intolerable. It is concluded that bortezomib at the dose of 1.0 mg/m² is recommended in combination with doxorubicin and intermediate-dose dexamethasone.

Allogeneic hematopoietic stem cell transplantation (HSCT) is increasingly used as a curative option for adult T-cell leukemia (ATL), an intractable mature T-cell neoplasm causally linked with human T-cell leukemia virus type I (HTLV-I). We compared outcomes of 386 patients with ATL who underwent allogeneic HSCT using different graft sources: 154 received human leukocyte antigen (HLA)–matched related marrow or peripheral blood; 43 received HLA-mismatched related marrow or peripheral blood; 99 received unrelated marrow; 90 received single unit unrelated cord blood. After a median follow-up of 41 months (range, 1.5-102), 3-year overall survival for entire cohort was 33% (95% confidence interval, 28%-38%). Multivariable analysis revealed 4 recipient factors significantly associated with lower survival rates: older age (> 50 years), male sex, status other than complete remission, and use of unrelated cord blood compared with use of HLA-matched related grafts. Treatment-related mortality rate was higher among patients given cord blood transplants; disease-associated mortality was higher among male recipients or those given transplants not in remission. Among patients who received related transplants, donor HTLV-I seropositivity adversely affected disease-associated mortality. In conclusion, allogeneic HSCT using currently available graft source is an effective treatment in selected patients with ATL, although greater effort is warranted to reduce treatment-related mortality.

The acquired JAK2 V617F mutation is observed in the majority of patients with BCR-ABL1 negative chronic myeloproliferative neoplasms (MPN). BCR-ABL1 negative MPN displays myeloproliferation with an elevated leucocyte alkaline phosphatase (LAP) activity, a neutrophil activation marker. We tried to separate the downstream signalling of JAK2 V617F to stimulate myeloproliferation and LAP activity. NB4, a myeloid lineage cell line, was transduced with Jak2 V617F mutation or wild-type Jak2. We found that Jak2 V617F mutation, but not wild-type Jak2 enhanced LAP expression in NB4-derived neutrophils and proliferation of NB4 cells. JAK2 V617F induces constitutive phosphorylation of STAT3 and STAT5, and uses signalling targets such as Ras/MEK/ERK and PI3K/Akt pathways. By using MEK1/2 inhibitor U0126, PI3K inhibitor LY294002, and STAT3 or STAT5 siRNAs, JAK2 V617F was found to specifically use the STAT3 pathway to enhance LAP expression, while STAT5, Ras/MEK/ERK and PI3K/Akt, but not STAT3 pathways, were able to stimulate cell proliferation. These data strongly suggest that JAK2 V617F uses distinct signalling pathways to induce typical pathological features of MPN, such as high LAP activity and enhanced cell proliferation.

Polyploid chromosomes are those with more than two sets of homologous chromosomes. Polyploid chromosomal abnormalities are observed in various malignant tumors. The prognosis in such cases is generally poor. However, there are no studies examining the prognosis of diffuse large B-cell lymphoma (DLBCL) with polyploid chromosomal abnormalities. Therefore, we statistically compared the clinicopathological features between polyploid DLBCL and DLBCL without polyploid abnormalities. Herein, 51 polyploid DLBCL and 53 control (without polyploid chromosomal abnormalities) cases were examined. G-banding method was employed to define polyploidy by cytogenetic analysis. Subsequently, flow cytometric immunophenotyping and immunohistochemical staining were performed. Polyploid DLBCL was defined as DLBCL with either near-tetraploid or greater number of chromosomes, as detected by the G-band. In a survival analysis, a significantly worse overall survival (OS) was observed for polyploid DLBCL (p = 0.04; p = 0.02 in cases who received R-CHOP regimens). In a multivariate analysis of OS, polyploid chromosomal abnormalities were an independent prognostic factor. Our results suggest that polyploid chromosomal abnormalities detected through G-band may represent a new poor prognostic factor for DLBCL.

Invasive aspergillosis (IA) is an important cause of infectious morbidity and mortality in patients who undergo haematopoietic stem cell transplantation (HSCT). History of IA before allogeneic HSCT is still challenging because of the high risk of recurrence after HSCT. Recent advances in early-stage diagnosis and new, more effective classes of antifungal agents have improved the management of IA in the HSCT recipients. We report two cases with acute myelogenous leukaemia after primary failure of induction chemotherapy with the patients developing pulmonary IA. They responded well to a combination of voriconazole (VCZ) and micafungin, resulting in a remarkable reduction of pulmonary IA lesions at short intervals. Thereafter, antifungal therapy was switched to liposomal amphotericin B (L-AmB), followed by conditioning regimen for allogeneic HSCT, because of the possibility of VCZ altering the metabolism of chemotherapeutic agents and calcineurin inhibitors. Successful engraftment was achieved without severe adverse side-effects or aggravation of IA after HSCT. Combining VCZ with micafungin followed by L-AmB throughout HSCT could be advantageous in stabilising IA in HSCT patients.

We performed a retrospective analysis of patients with diffuse large B cell lymphoma treated with rituximab plus CHOP (cyclophosphamide, adriamycin, vincristine and prednisone) as a first-line therapy at 22 hospitals of the Kyushu Lymphoma Study Group. During the period 1996-2005, 1,057 patients (aged 22-90 years) were analyzed. Of these, 678 were treated with CHOP, and 379 were treated with rituximab plus CHOP (R-CHOP). The complete response rate was 59.9% in the CHOP group and 67.0% in the R-CHOP group (P0.001). Three-year progression-free survival (PFS) and overall survival (OS) rates were significantly higher in the R-CHOP group than in the CHOP group (61.3 vs. 45.6% for PFS, P0.001; 68.3 vs. 54.5% for OS, P0.001). The International Prognostic Index was a good prognostic marker for both groups; a survival benefit of rituximab addition was found for each risk subgroup and also for both age groups (or=60 and60 years). Among 345 patients who received localized radiation therapy, the adding rituximab to CHOP attenuated the survival difference between CHOP and R-CHOP groups (P = 0.104), compared with no radiation group (P0.001). Results of this large-scale, multicenter study confirm that rituximab plus CHOP provided a greater survival benefit than CHOP alone.

Invasive aspergillosis (IA) is a major cause of morbidity and mortality among the immunocompromised, especially those undergoing hematopoietic stem cell transplantation. With spore inhalation the usual infection route, such subjects must be protected from environmental spore contamination, necessitating measures such as high-efficiency particulate air (HEPA) filtration. In April 2006, we implemented a new transplantation unit with HEPA filtration. We retrospectively evaluated its efficacy for hospitalized transplantation unit subjects whose sera were tested for aspergillus galactomannan antigen between April 2004 and March 2007. Subjects numbered 265 (973 samples) categorized as definite, probable, or possible. The earliest IA onset date was when symptoms, positive radiological findings, or positive galactomannan antigen tests occurred, based on revised European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) definitions. We classified cases when IA occurred over 10 days after admission as hospital-acquired. No such cases were detected after November 2005 and IA incidence decreased significantly after the new unit began being used. Results suggest that the new unit and HEPA filtration helped eliminate nosocomial IA.

We prospectively evaluated the safety and efficacy of the anti-CD20 chimeric monoclonal antibody rituximab for the treatment of corticosteroid-refractory chronic graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation. Seven patients were treated with 375 mg/m(2) rituximab weekly for 4 consecutive weeks. Rituximab was well tolerated with no severe toxicity observed during treatment. At 1 year, 3 patients showed a partial response to rituximab therapy, 3 had stable disease, and 1 had progressive disease. Rituximab allowed a reduction in the dose of steroids in 4 patients. Responsive manifestations included mild to moderate skin and oral lesions, and immune hemolytic anemia, and thrombocytopenia. Severe manifestations involving the skin, fascia, and eye did not respond to treatment. These observations suggest that rituximab therapy may be effective for select patients with corticosteroid-refractory chronic GVHD that is not advanced.

We describe a case of encephalomyelitis mimicking multiple sclerosis associated with chronic graft-versus-host disease (GVHD) occurring after allogeneic bone marrow transplantation (BMT) for myelodysplastic syndrome. Immunosuppressive therapy, consisting of a therapeutic dose of cyclosporine A and a maintenance dose of methylprednisolone, was effective in treating symptoms. Although central nervous system GVHD is very rare and remains controversial, presentation of neurological symptoms after allogeneic BMT warrants consideration of GVHD in the differential diagnosis.

To establish effective therapeutic strategies for eosinophil-related disorders, it is critical to understand the developmental pathway of human eosinophils. In mouse hematopoiesis, eosinophils originate from the eosinophil lineage-committed progenitor (EoP) that has been purified downstream of the granulocyte/macrophage progenitor (GMP). We show that the EoP is also isolatable in human adult bone marrow. The previously defined human common myeloid progenitor (hCMP) population (Manz, M.G., T. Miyamoto, K. Akashi, and I.L. Weissman. 2002. Proc. Natl. Acad. Sci. USA. 99:11872-11877) was composed of the interleukin 5 receptor alpha chain(+) (IL-5Ralpha(+)) and IL-5Ralpha(-) fractions, and the former was the hEoP. The IL-5Ralpha(+)CD34(+)CD38(+)IL-3Ralpha(+)CD45RA(-) hEoPs gave rise exclusively to pure eosinophil colonies but never differentiated into basophils or neutrophils. The IL-5Ralpha(-) hCMP generated the hEoP together with the hGMP or the human megakaryocyte/erythrocyte progenitor (hMEP), whereas hGMPs or hMEPs never differentiated into eosinophils. Importantly, the number of hEoPs increased up to 20% of the conventional hCMP population in the bone marrow of patients with eosinophilia, suggesting that the hEoP stage is involved in eosinophil differentiation and expansion in vivo. Accordingly, the phenotypic definition of hCMP should be revised to exclude the hEoP; an "IL-5Ralpha-negative" criterion should be added to define more homogenous hCMP. The newly identified hEoP is a powerful tool in studying pathogenesis of eosinophilia and could be a therapeutic target for a variety of eosinophil-related disorders.

We report a case of severe parainfluenza (PIV) 3 pneumonia in a hematopoietic stem cell transplant recipient that was successfully treated with oral ribavirin and methylprednisolone. A 42-year-old woman diagnosed with acute myelogenous leukemia (FAB M5a) in first complete remission underwent allogeneic bone marrow transplantation from an HLA-matched unrelated donor in May 2006. In July 2007, she developed PIV3 pneumonia. Her respiratory status progressively worsened and she required O(2) inhalation at 6 L/min. After an informed consent was obtained, oral ribavirin was initiated (16 mg/kg per day) for 1 week on July 31. By day 3 of treatment, the high-grade fever had disappeared. However, it recurred after ribavirin was discontinued. In addition, the patient's hypoxia continued to worsen, requiring O(2) inhalation at 9 L/min. To suppress the inflammatory reaction in the lung caused by PIV3 pneumonia, intravenous methylprednisolone (1,000 mg once a day for 3 days) was started along with high-dose oral ribavirin (16 mg/kg per day) on August 11. The patient showed dramatic clinical improvement, and oxygen inhalation was discontinued on September 3. Our case suggests that with concomitant effective anti-viral treatment, corticosteroids may suppress host inflammatory or immune reactions that lead to respiratory failure.

FLT3/FLK2, a member of the receptor tyrosine kinase family, plays a critical role in maintenance of hematopoietic homeostasis, and the constitutively active form of the FLT3 mutation is one of the most common genetic abnormalities in acute myelogenous leukemia. In murine hematopoiesis, Flt3 is not expressed in self-renewing hematopoietic stem cells, but its expression is restricted to the multipotent and the lymphoid progenitor stages at which cells are incapable of self-renewal. We extensively analyzed the expression of Flt3 in human (h) hematopoiesis. Strikingly, in both the bone marrow and the cord blood, the human hematopoietic stem cell population capable of long-term reconstitution in xenogeneic hosts uniformly expressed Flt3. Furthermore, human Flt3 is expressed not only in early lymphoid progenitors, but also in progenitors continuously along the granulocyte/macrophage pathway, including the common myeloid progenitor and the granulocyte/macrophage progenitor. We further found that human Flt3 signaling prevents stem and progenitors from spontaneous apoptotic cell death at least through up-regulating Mcl-1, an indispensable survival factor for hematopoiesis. Thus, the distribution of Flt3 expression is considerably different in human and mouse hematopoiesis, and human FLT3 signaling might play an important role in cell survival, especially at stem and progenitor cells that are critical cellular targets for acute myelogenous leukemia transformation.

A major issue in autologous blood stem cell transplantation (ABSCT) for leukaemia is whether peripheral blood stem cell (PBSC) harvests are less contaminated with leukaemic cells than bone marrow mononuclear cells (BMMNC). We compared leukaemic contamination in PBSC harvests and BMMNC, obtained simultaneously, by using reverse transcriptase polymerase chain reaction (RT-PCR) of leukaemia-specific chimaeric messenger RNA (mRNA), in three patients with Philadelphia chromosome (Ph)-positive acute lymphoblastic leukaemia (ALL), one with Ph-positive acute myelogenous leukaemia (AML), and two with acute promyelocytic leukaemia (APL). Our two-step PCR method employed 'nested primers' in the second step and can detect one leukaemic blast diluted into 10(6) HL-60 cells. In three of four patients with Ph-positive ALL and AML we detected leukaemic contamination in both PBSC harvests and BMMNC. In the remaining patient with ALL, both PBSC harvests and BMMNC were PCR-negative. Both PBSC harvests and BMMNC from one patient with APL were PCR-positive. In contrast, PBSC harvests from another patient with APL, whose BMMNC could not be obtained because of bone marrow necrosis, were PCR-positive after the first course of consolidation chemotherapy, but became PCR-negative after the second course. The present study does not support the hypothesis that PBSC harvests are less contaminated by leukaemic cells than BMMNC, but suggests that PBSC harvests are contaminated when BMMNC are contaminated.

Primary plasma cell leukaemia (PCL) is a rare, aggressive neoplasm of plasma cell dyscrasia. Conventional chemotherapy is usually ineffective, with an overall survival of only 8 months. Here, we describe a 42-year-old man with primary PCL, who was successfully treated with haploidentical (2-HLA loci mismatched) haematopoietic stem-cell transplantation (HSCT). To overcome the human leukocyte antigen (HLA) disparity, in vivo T-cell purging by the pre-transplant administration of antithymocyte globulin followed by a conventional prophylactic treatment against graft-versus-host disease (GVHD) resulted in an avoidance of severe GVHD as well as infectious complications. The patient has maintained complete remission for 13 months after haploidentical HSCT, indicating that a graft-versus-PCL effect might be preserved. Haploidentical HSCT can be a potentially curative treatment for patients with primary PCL who do not have an HLA-identical donor.

Toxoplasmosis is a rare but rapidly fatal complication that can occur following hematopoietic stem cell transplantation (HSCT). Over a 17-yr period at our institutions, a definite diagnosis of toxoplasmosis was made in only two of 925 allogeneic HSCT recipients (0.22%) and none of 641 autologous HSCT recipients. These two patients received a conventional conditioning regimen followed by transplantation from an HLA-matched donor; however, they developed severe graft-vs.-host disease, which required intensive immunosuppressive therapy. Despite prophylactic treatment with trimethoprim/sulfamethoxazole, their immunosuppressive state, as indicated by a low CD4(+) cell count, might have resulted in toxoplasmosis encephalitis. Rapid and non-invasive methods such as a polymerase chain reaction (PCR) test of their cerebrospinal fluid for Toxoplasma gondii and magnetic resonance imaging of the brain were useful for providing a definitive diagnosis and prompt therapy in these patients: one patient stabilized and survived after responding to treatment with pyrimethamine/sulfodiazine whereas the other died of bacterial infection. In addition, retrospective PCR analyses of the frozen stored peripheral blood samples disclosed that detection of T. gondii preceded the onset of disease, indicating routine PCR testing of peripheral blood specimens may be an early diagnostic tool. It should be noted that when patients receiving HSCT have an unexplained fever and/or neurological complications, PCR tests should be considered to avoid cerebral lesions and improve the outcome of the patients.

Objective: Patients with acute myelogenous leukaemia (AML) show co-existing frequently internal tandem duplications of FLT3 (FLT3-ITD) and mutations of nucleophosmin (NPM1-Mt). We investigated the biological and clinical significance of FLT3-ITD and/or NPM1-Mt in this context. Methods: We analysed 89 AML patients according to whether NPM1 and FLT3-ITD were single mutants, double mutants, or wild type for both. Results: FLT3-ITD was detected in 19 of 89 patients (21.3%), while NPM1-Mt was detected in 19 of 89 patients (21.3%); eight of 89 patients (9.0%) carried both FLT3-ITD and NPM1-Mt. By multivariate analysis, white blood cell count and peripheral blood blast cell count at diagnosis were significantly higher in patients with FLT3-ITD but not in those with only NPM1-Mt. NPM1-Mt was significantly related to female gender, normal karyotype, and M4 or M5 disease according to French–American–British criteria. In addition, leukaemic blast cells with NPM1-Mt, FLT3-ITD, or both expressed CD34 less frequently than wild-type blasts (P

Maintaining the appropriate average steady-state plasma concentrations (Css) of busulfan (BU) is critical for both successful engraftment and minimizing toxicity in hematopoietic stem cell transplantation (HST). We therefore performed a prospective trial with 50 adult Japanese patients that involved adjusting the BU dose in accordance with individual BU pharmacokinetics (PK). After administering a 0.5-mg/kg test dose of oral BU, we analyzed individual BU PK parameters and calculated an adjusted BU dose that would achieve a target BU Css of 850 ng/mL. Thirty-nine patients (78%) required a BU dose decrease, and the median adjusted BU dose was 0.81 mg/kg (range, 0.51-1.29 mg/kg). All patients who underwent allogeneic HST received the adjusted BU dose. After administering the sixth BU dose, we measured the plasma BU concentration. The actual BU concentration was significantly correlated with the expected BU concentration, and the predictability of the BU Css was 103% +/- 9%. The incidence of toxicity excluding oral mucositis was low, and there was no regimen-related toxicity-associated mortality. Engraftment was achieved in 98% of the patients. This study showed that our method for adjusting the BU dose facilitated reliable prediction of the actual BU Css and that individualized BU dose adjustment was able to improve clinical outcomes in HST recipients treated with a BU-containing conditioning regimen.

Behçet's disease is a chronic, relapsing, inflammatory disease of unknown origin. The association of myelodysplastic syndrome and Behçet's disease is rare, and recent reports have indicated that immunosuppressive agents alone are not sufficient to control Behçet's disease associated with MDS and many patients die of infection or hemorrhage. We report a case of MDS with intestinal Behçet's disease. We performed cord blood transplantation with a myeloablative regimen as the primary treatment. The patient achieved complete remission for both diseases, which continued for more than 16 months. Our experience suggests that CBT may provide a potent therapeutic option for the treatment of MDS-related Behçet's disease.

Isolated extramedullary (EM) relapses of acute myelogenous leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) have been reported to be rare, and are usually followed by bone marrow relapses. We report a 49-year-old man with AML with the unfavorable chromosome abnormality 7q-, who was treated by allo-HSCT. Fifteen months after allo-HSCT, the patient initially developed a relapse only in his inguinal lymph nodes, and then bone marrow relapse became evident one month after the EM relapse. Subsequently, the patient received chemotherapy and a second allo-HSCT from another donor, but he suffered another relapse in different EM sites including the skin and central nervous system with a persistently normal marrow. This case is characterized by repeated relapses in isolated EM sites after allo-HSCT and suggests that the anti-leukemic effects of chemotherapy and/or graft-versus-leukemia effects in the EM sites might not be so uniformly effective as that in the marrow. Accordingly, we should be aware that AML relapses can occur repeatedly only in isolated EM sites post allo-HSCT, resulting in treatment failure and a poor prognosis.

To evaluate the efficacy and feasibility of upfront high-dose chemotherapy (HDCT) and rituximab (R) followed by autologous peripheral blood stem cell transplantation (auto-PBSCT) in patients with newly diagnosed high-intermediate(HI)-, and high(H)-risk diffuse large B-cell lymphoma (DLBCL), we conducted a multicenter prospective phase II trial. In 15–60-year-old patients with H- or HI-risk DLBCL, after three courses of (R-)CHOP14, high-dose etoposide was given prior to peripheral blood stem cell harvesting. After an additional three courses of (R-)CHOP14, auto-PBSCT was performed following HDCT. The primary endpoint of the study is progression-free survival (PFS) at 2 years after registration in eligible patients. The expected PFS and the threshold PFS were estimated to be 70 and 50 %, respectively. Among 40 eligible patients registered, 30 patients completed treatment. With a median observation period in surviving eligible patients of 63 months, the 2- and 4-year PFS after registration were 79.9 and 72.0 %, respectively. The 2- and 4-year overall survival (OS) were 92.5 and 84.6 %, respectively. In 30 patients who completed treatment, the 4-year PFS and OS after auto-PBSCT were 79.2 and 85.9 %, respectively. In conclusion, the results of our study suggest that upfront HDCT and auto-PBSCT combined with rituximab is highly effective as an initial treatment for HI-, and H-risk DLBCL.

Programmed cell death ligand 1 (PD-L1) is expressed on both select diffuse large B-cell lymphoma (DLBCL) tumor cells and on tumor-infiltrating nonmalignant cells. The programmed cell death 1 (PD-1)/PD-L1 pathway inhibits host antitumor responses; however, little is known about how this pathway functions in the tumor microenvironment. The aim of this study was to determine the clinicopathological impact of PD-L1(+) DLBCL. We performed PD-L1/PAX5 double immunostaining in 1253 DLBCL biopsy samples and established a new definition of PD-L1(+) DLBCL. We also defined the criteria for microenvironmental PD-L1(+) (mPD-L1(+)) DLBCL (ie, PD-L1(-) DLBCL in which PD-L1(+) nonmalignant cells are abundant in the tumor microenvironment). Of the 273 patients whose clinical information was available, quantitative analysis of PD-1(+) tumor-infiltrating lymphocytes (TILs) was performed. The prevalence rates of PD-L1(+) and mPD-L1(+) DLBCL were 11% and 15.3%, respectively. Both PD-L1(+) and mPD-L1(+) DLBCL were significantly associated with non-germinal center B-cell (GCB) type and Epstein-Barr virus positivity. The number of PD-1(+) TILs was significantly higher in GCB-type tumors and lower in mPD-L1(-) and PD-L1(+) DLBCL. Patients with PD-L1(+) DLBCL had inferior overall survival (OS) compared with that in patients with PD-L1(-) DLBCL (P = .0009). In contrast, there was no significant difference in OS between mPD-L1(+) and mPD-L1(-) DLBCL (P = .31). The expression of PD-L1 maintained prognostic value for OS in multivariate analysis (P = .0323). This is the first report describing the clinicopathological features and outcomes of PD-L1(+) DLBCL. Immunotherapy targeting the PD-1/PD-L1 pathway should be considered in this distinct DLBCL subgroup.

血液型不適合骨髄移植を行う際は, 輸注に伴う有害事象を避けるため, 骨髄液中の赤血球・血漿除去が必要である. 今回われわれは, 血液成分分離装置COBE Spectra により, 骨髄液から単核球を分離することで, 骨髄細胞濃縮を行った血液型不適合骨髄移植20例において, その有用性について検討した. COBE Spectra を用いた骨髄濃縮により, 赤血球量は98.4%除去され, 最終赤血球量は4.2±2.4mlであった. 有核細胞回収率は34.0±8.38%, CD34陽性細胞回収率は112.3±36.3%であった. 20症例全例で, 移植後の溶血反応および生着不全は認めず, 移植後造血回復は速やかであった. COBE Spectra を用いることで, 清潔な無菌閉鎖回路内において骨髄濃縮が可能となり, 処理時間も短縮された. 種々の細胞免疫療法の用途で広く普及しているCOBE Spectra を用いた骨髄濃縮法は, 血液型不適合骨髄移植における骨髄濃縮の標準的方法として有用であると考えられた.

Hemopoietic stem and progenitor cells ordinarily residing within bone marrow are released into the circulation following G-CSF administration. Such mobilization has a great clinical impact on hemopoietic stem cell transplantation. Underlying mechanisms are incompletely understood, but may involve G-CSF-induced modulation of chemokines, adhesion molecules, and proteolytic enzymes. We studied G-CSF-induced mobilization of CD34+CD10+CD19−Lin− and CD34+CD10+CD19+Lin− cells (early B and pro-B cells, respectively). These mobilized lymphoid populations could differentiate only into B/NK cells or B cells equivalent to their marrow counterparts. Mobilized lymphoid progenitors expressed lymphoid- but not myeloid-related genes including the G-CSF receptor gene, and displayed the same pattern of Ig rearrangement status as their bone marrow counterparts. Decreased expression of VLA-4 and CXCR-4 on mobilized lymphoid progenitors as well as multipotent and myeloid progenitors indicated lineage-independent involvement of these molecules in G-CSF-induced mobilization. The results suggest that by acting through multiple trans-acting signals, G-CSF can mobilize not only myeloid-committed populations but a variety of resident marrow cell populations including lymphoid progenitors.

A patient with intracerebral hemorrhage is considered ineligible for hematopoietic stem cell transplantation (HSCT). We report a 49-year-old woman with myelodysplastic syndrome (MDS) complicated by refractoriness to platelet transfusion and intracerebral hemorrhage, who underwent allogeneic bone marrow transplantation from an HLA-identical unrelated male donor. Nine days before the scheduled transplantation, she developed dysarthria and right hemiparesis; computed tomography (CT) of the brain disclosed an acute hematoma in the left parietal lobe exceeding 3 cm in diameter. She underwent conditioning with reduced-intensity, including fludarabine (30 mg/m2/day on days -8 to -3), busulfan (4 mg/kg/day on days -6 and -5), and total body irradiation (4 Gy on day -2). Two weeks after transplantation, dysarthria and right hemiparesis began to resolve, and CT showed spontaneous resolution of the hematoma. Simultaneously, engraftment was confirmed. Thus, recent stroke may be not an absolute contraindication for HSCT.

We report a 72-year-old man with Waldenström's macroglobulinemia (WM) in whom diffuse large B-cell lymphoma (DLCL) occurred 17 years after the diagnosis of WM. The malignant cells of both DLCL and WM expressed CD20 on their surface. CHOP plus anti-CD20 monoclonal antibody, rituximab, were effective for both diseases, and the patient remains disease-free 17 months later.

We report herein a 19-year-old Japanese male with XYY syndrome who developed acute myelogenous leukemia. During three courses of cytotoxic chemotherapy, he suffered repeated hepatic and renal insufficiencies, possibly related to latent dysfunction from the XYY syndrome. The patient was treated with granulocyte colony-stimulating factor combined with etoposide, cytarabine, and busulfan (the latter adjusted to a targeting dose) followed by autologous peripheral blood stem cell transplantation. He had no severe regimen-related toxicities and is now free of leukemia.

Objective Limitin, an interferon-like cytokine, suppresses B lymphopoiesis through ligation of the interferon-α/β (IFN-α/β) receptor. The aim of this study was to examine the intracellular signal transduction pathways activated by limitin. Materials and methods The effects of limitin on cell growth, the activation of Jak kinase and Stat proteins, and the induction of interferon regulatory factor-1 (IRF-1) and Daxx were examined using the mouse pre–B-cell line 18.81, wild-type, and Tyk2-deficient mouse bone marrow cells. In addition, the change of localization of the Daxx protein after limitin treatment in wild-type and Tyk2-deficient mice was examined. Results Limitin phosphorylates Tyk2, Jak1, Stat1, and Stat2 and rapidly induces IRF-1 mRNA production. Phosphorylation of Stat1 by limitin is partially dependent on Tyk2. Suppression of B-cell growth by limitin, however, is severely impaired in the absence of Tyk2, whereas it is unaffected by the absence of Stat1. Limitin also induces the expression and nuclear translocation of Daxx, which is essential for IFN-α–induced inhibition of B-lymphocyte development. The absence of Tyk2 abrogates this induction of Daxx expression and nuclear translocation. Conclusions Limitin suppresses B-cell growth through activation of Tyk2, resulting in the up-regulation and nuclear translocation of Daxx. This limitin-mediated signaling pathway does not require Stat1.

Many hematopoietic cells undergo apoptosis when deprived of specific cytokines. Lipocalin 24p3, reported to be induced in hematopoietic cells by interleukin 3 (IL-3) depletion, induces hematopoietic cell apoptosis despite the presence of IL-3. As granulocyte colony stimulating factor (G-CSF) depletion also induces the apoptosis of G-CSF-dependent cell line cells, we examined the effect of 24p3 on the apoptotic function induced by G-CSF depletion. 24p3 was induced by the depletion of IL-3, but not G-CSF, in cytokine-dependent cell lines. Although 24p3 suppressed growth induced by IL-3, it did not influence G-CSF-dependent cell growth. These observations show that 24p3 is not involved in the G-CSF withdrawal-induced apoptosis, although it is essential in IL-3 withdrawal-induced apoptosis.

Interferon (IFN)-alpha and IFN-gamma suppress the growth of haematopoietic progenitor cells. IFN-alpha activates Janus kinase-1 (Jak1) and Tyrosine kinase-2 (Tyk2), followed by the phosphorylation of the signal transducers and activators of transcription, Stat1 and Stat2. IFN-gamma activates Jak1 and Jak2, followed by the activation of Stat1. Activated Stats bind the promoter regions of IFN-inducible genes. We evaluated the role of Tyk2 and Stat1 in the IFN-mediated inhibition of haematopoietic progenitor cell growth. While IFN-alpha (1000 U/ml) suppressed the number of granulocyte-macrophage colony-forming units (CFU-GM) or erythroid burst-forming units (BFU-E) from wild-type mouse bone marrow cells, this suppression was partially inhibited by a deficiency in Tyk2 and completely inhibited by a deficiency in Stat1. High levels of IFN-alpha (10,000 U/ml) suppressed the CFU-GM or BFU-E obtained from Stat1-deficient mice, but did not suppress this growth in cells from Tyk2-deficient mice. Stat1 was phosphorylated by IFN-alpha in Tyk2-deficient cells, although the level of phosphorylation was weaker than that observed in wild type mice. Thus, the inhibitory signal on haematopoietic progenitor cells mediated by IFN-alpha may be transduced by two signalling pathways, one regulated by Tyk2 and the other dependent on Stat1. IFN-gamma also suppressed the number of CFU-GM or BFU-E, and this pathway was mediated by IFN-gamma in a Stat1-dependent manner, independently of Tyk2.

We investigated effects of variations in the cellular composition of G-CSF-mobilized peripheral blood progenitor cell (G-PBPC) allografts on clinical outcomes of allogeneic PBPC transplantation. We retrospectively analyzed transplanted doses of various immunocompetent cells from 27 HLA-identical sibling donors in relation to engraftment, incidence of graft-versus-host disease (GVHD), and survival. Significant variability was documented in both absolute numbers and relative proportions of CD34+, CD2+, CD3+, CD4(high)+, CD4+25+, CD8(high)+, CD19+, CD56+, and CD56+16+ cells contained in these allografts. Stepwise Cox regression analysis revealed that the CD56+ cell dose was significantly inversely correlated with the incidence of GVHD. Thus, there was a significantly higher incidence of grade II acute GVHD in patients receiving a lower CD56+16+ cell dose (hazard ratio (HR) 0.0090; 95% confidence interval (CI)

A 74-year-old man was referred to us for evaluation of a tumor in the right atrium (RA). Transesophageal echocardiography (TEE) showed an unmovable 50x60 mm mass in the RA. Based on histological findings of subcutaneous tumors in the right abdominal wall, he was diagnosed as malignant lymphoma (ML), and treated with a THP-COP regimen. Upon completion of first THO-COP therapy, TEE showed marked regression of the mass and division into 3 masses, one of which showed marked floating movement with a small stalk. To prevent the risk of embolic events, surgical resection was performed. Resected tumors were necrotic tissues. Serial imaging of cardiac tumor and surgical resection is desirable to decrease the possibility of embolic complication.

Viral hemorrhagic cystitis (HC) is a frequent complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) but is rare after autologous peripheral blood stem cell transplantation (auto-PBSCT) because immunosuppression is not required in the absence of an allogeneic immune reaction. Recently, auto-PBSCT has been combined with novel anticancer agents targeting specific molecules, such as rituximab; however, these may cause severe immune deficiency and increase the susceptibility of transplant recipients to opportunistic infections. To address this issue, we performed a retrospective analysis of the incidence of viral HC in auto-PBSCT recipients. Of 158 recipients, only 4 cases (2.5%) were diagnosed with viral HC due to adenovirus (ADV), which was significantly less frequent compared with the incidence in allo-HSCT recipients (15.8%). The incidence of HC did not increase with rituximab treatment. This was a single-center retrospective analysis with a small sample size; however, incorporation of rituximab into the treatment of auto-PBSCT recipients did not appear to be a risk factor for developing viral HC. Viral HC is a noteworthy complication after HSCT because of its negative effects on the quality of life and its potential to cause therapy-related mortality [1]. We recently reported different risk factors associated with two viral HCs in allo-HSCT recipients [2]: ADV-HC and BKV-HC. ADV-HC is associated with severe immunodeficiency caused by immunosuppression therapies such as T-cell purging, whereas BKV-HC is associated with allogeneic immune hyperactivity such as graft-versus-host disease (GVHD) or the pre-engraftment immune reaction (PIR) [3]. Auto-PBSCT is characterized by rapid hematologic and immune reconstitution and does not trigger an allogeneic immune reaction that would require immunosuppressive therapy either for prophylaxis or for treatment. Consequently, there is lower incidence and severity of opportunistic infection including viral HC in auto-PBSCT recipients [4,5]. The survival of auto-PBSCT recipients has significantly improved over the recent years because of the incorporation of novel anticancer agents targeting specific molecules into the treatment regimen [6]. One such anticancer agent widely used in this context is rituximab, a chimeric monoclonal anti-CD20 antibody that causes B-lymphocyte depletion. When used alone in patients with lymphoma, rituximab has few effects on the incidence of infectious complications [7,8]. However, the combination of rituximab and high-dose chemotherapy (HDC) may increase the risk of viral infections [9–11], including that caused by the hepatitis-B virus (HBV) [12]. This retrospective study was conducted to clarify the prevalence, clinical features, and risk factors of viral HC after auto-PBSCTand to compare these variables with those of allo-HSCT. A total of 158 patients with hematological disorders or solid tumors hospitalized at our institute from 2000 to 2010 were enrolled. The clinical characteristics of these patients are summarized in Table I. A total of 101 patients had malignant lymphoma (ML), and the remaining 57 patients were diagnosed with acute myelogenous leukemia (AML; n 5 13), plasma-cell disorders (e.g., multiple myeloma or primary amyloidosis; n 5 36), or solid tumors (n 5 8). Patients with one of the following conditions were classified as ‘‘chemotherapy-sensitive’’ (n 5 132): ML and plasma cell disorders in at least partial remission and AML in first complete remission. All other patients (n 5 26) were categorized as ‘‘chemotherapy-resistant.’’ A total of 101 ML patients were treated with the CHOP regimen [13] as induction therapy, HDC as a preparative conditioning regimen, followed by transplantation of unpurged autologous PBSCs. After approval of rituximab for the treatment of ML in Japan, 41 patients with CD20-positive lymphomas was treated with CHOP and auto-PBSCT combined with rituximab. Of these, 35 patients received rituximab in combination with the ranimustine, carboplatin, etoposide, and cyclophosphamide (MCEC) pretransplant regimen [14]. Four patients received rituximab combined with the melphalan, etoposide, cyclophosphamide, and dexamethasone (LEED) regimen [15] and 2 patients received rituximab in combination with other regimens. The remaining 60 ML patients were either treated before approval of the use of rituximab or had CD20-negative lymphoma and received HDC consisting of MCEC (n 5 52), LEED (n 5 2), or another regimen (n 5 6), all without rituximab. Plasma cell disorders (n 5 36) were treated with the VAD regimen [16] or bortezomib-based induction therapy followed by high-dose melphalan before auto-PBSCT [17]. Standard-risk AML patients (n 5 13) achieved complete remission after conventional remission induction and consolidation chemotherapy and received the busulfan, cytarabine, and etoposide (BEA) conditioning regimen followed by auto-PBSCT [18]. Eight patients with solid tumors underwent standard chemotherapies based on each diagnosis and then received auto-PBSCT. Of these, five patients received the high-dose ICE regimen [19]. Acyclovir prophylaxis was used to protect against reactivation of herpes simplex virus (HSV) in all these recipients. Urinalysis was routinely performed at least once a week from the initiation of preparative regimens until discharge from hospital or when clinical signs of cystitis appeared after the discharge. Microscopic or macroscopic hematuria and/or bladder irritation was further analyzed using polymerase chain reaction (PCR) to detect viral DNA (ADV, BKV, and JC virus). Methods for detecting each type of viral DNA have been reported previously [20–22]. In brief, after 2 ml of a urine sample was centrifuged (15,000g for 1 hr), DNA was purified from the sediment using the QIAmp DNA Blood Mini Kit (Qiagen, Hilden, Germany). Next, 5 ml of purified DNA was subjected to PCR using the GeneAmp Kit and GeneAmp PCR System 9600 (PerkinElmer, Boston, MA) with the following primers: AD185S (50-tccagcaacttcatgtccatgg-30) and AD 185A (50-tcgatgacgccgcggt30) for detecting ADV; BKV-1 (50-gcaagtgccaaaactactaat-30) and BKV-2 (50tgcatgaaggttaagcatgc-30) for detecting BKV; and JTP-1 (50-gcagcttagtgattttct

IFN-α inhibits B lymphocyte development, and the nuclear protein Daxx has been reported to be essential for this biological activity. We show in this study that IFN-α inhibits the clonal proliferation of B lymphocyte progenitors in response to IL-7 in wild-type, but not in tyk2-deficient, mice. In addition, the IFN-α-induced up-regulation and nuclear translocation of Daxx are completely abrogated in the absence of tyk2. Therefore, tyk2 is directly involved in IFN-α signaling for the induction and translocation of Daxx, which may result in B lymphocyte growth arrest and/or apoptosis.

Introduction Widespread use of tyrosine kinase inhibitors (TKIs) in combination with chemotherapy and allogeneic hematopoietic stem cell transplantation (allo-SCT) has totally changed the existing treatment strategies for Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph + ALL). However, the prognosis after relapse after allo-SCT is still dismal. Patients and Methods We analyzed the clinical outcome of therapy using dasatinib, a second-generation TKI, in 9 patients with relapsed Ph + ALL after allo-SCT. Dasatinib was initiated at a median time of 168 days after allo-SCT at dosages ranging from 20 mg to 100 mg daily. Results Six of 9 patients manifested a marked increase in large granular lymphocytes (LGLs), but all 6 patients discontinued dasatinib because of adverse events (AEs) such as pleural effusion. Four of 6 patients resumed dasatinib, and 3 of them have been alive with molecular complete remission and a persistent increase of LGLs. Conclusion Our results demonstrated that dasatinib therapy can induce LGL expansion accompanied by AEs, but this phenomenon can be associated with long-term survival benefit in a proportion of relapsed Ph + ALL patients after allo-SCT.