Mayumi Hatsuse

A newly developed bisphosphonate, YM529, is a potent apoptosis inducer of human myeloma cells.

We examined the effect of YM529, a newly developed third-generation bisphosphonate (BP), on the growth of human myeloma cell lines using the trypan blue dye exclusion test and Alamar blue assay. BPs induced inhibition of proliferation in all cell lines dose-dependently, and YM529 had a most potent growth inhibitory effect, followed by incadronate and pamidronate. Flow cytometric analysis using annexinV and 7AAD showed that YM529 most significantly induced apoptosis of all myeloma cell lines. These observations suggested that YM529 is a potent apoptosis inducer of myeloma cells, and might have some benefit not only on the improvement of bone lesions but also on survival in some myeloma patients.

Expression of T‐cell‐associated antigens in B‐cell non‐Hodgkin's lymphoma

We performed the immunophenotyping of 101 patients with B‐cell non‐Hodgkins lymphoma (B‐NHL) using two‐colour flow cytometry (FCM) and found that lymphoma cells coexpressed at least one kind of T‐cell‐associated antigen (T‐Ag; CD2, CD5, CD7) in 25 patients (24·8%). Among these three T‐Ags, CD5 was the most frequently expressed, in 21 patients (20·8%), followed by CD7, expressed in five patients (5·0%), and CD2, which was expressed in two patients (2·0%). Two kinds of T‐Ag were simultaneusly expressed in three patients (CD2/CD5, CD2/CD7, and CD5/CD7, each expressed in one patient). Concerning the expression pattern of T‐Ag, there were no significant differences between lymph nodes and extranodal organs in the three patients with T‐Ag‐positive B‐NHL (T‐Ag(+) B‐NHL) who were analysed. When comparing the clinical features between T‐Ag(+) B‐NHL and T‐Ag‐negative B‐NHL (T‐Ag(–) B‐NHL), extranodal involvement and higher International Prognostic Index (H and H.I.) were significantly frequent in the former subgroup (P = 0·0119 and P = 0·0302 respectively).

CD34 + /CD90 + cells infused best predict late haematopoietic reconstitution following autologous peripheral blood stem cell transplantation

Summary.  This study aimed to identify which graft product subset of cells might be the most predictive of late haematopoietic recovery (three to 12 months) following autologous peripheral blood stem cell transplantation (PBSCT). The relationships between the numbers of reinfused CD34+ cells and their immature subsets such as CD34+/CD90+, CD34+/AC133+, CD34+/CD38– and CD34+/HLA‐DR– cells, and haemoglobin, white blood cell (WBC) and platelet counts at 3, 6, 9 and 12 months after PBSCT, were studied in 25 patients with haematological and solid malignancies. The total CD34+ cell number, as well as CD34+/CD90+ and CD34+/AC133+ cell numbers, correlated with platelet counts at 3, 6, 9 and 12 months after PBSCT, but the CD34+/CD90+ cells infused best predicted platelet recovery during the first 12 months after PBSCT (P < 0·0238 at any time‐point). The CD34+/AC133+ cell dose also correlated with WBC counts at 3 months post PBSCT. In addition, all patients receiving more than 80 × 104 CD34+/CD90+ cells/kg showed platelet counts greater than 100 × 109/l at all points after PBSCT, suggesting that this value of the CD34+/CD90+ cells infused was a threshold dose for durable haematopoietic engraftment after PBSCT.

Clinicopathological features of myeloid/natural killer (NK) cell precursor acute leukemia

Four patients (three males and one female) were diagnosed as myeloid/natural killer (NK) cell precursor acute leukemia in our department. Two patients showed the extramedullary involvement at initial presentation. Leukemic cells expressed CD7, CD33, CD45 and CD56 in all patients. Additionally, CD13, CD34, HLA-DR, cytoplasmic CD3 and myeloperoxidase were expressed in some patients. Trisomy 10 was found in two patients, which has not been reported in this disease. Therefore, myeloid/NK cell precursor acute leukemia might be rather heterogeneous especially in chromosomal abnormality though it seemed to constitute the distinct clinical entity among acute myeloid leukemia of M0 subtype.

Combination Therapy with Thalidomide, Incadronate, and Dexamethasone for Relapsed or Refractory Multiple Myeloma

The feasibility and efficacy of a combination of thalidomide, incadronate, and dexamethasone (TID) were studied in 12 patients with relapsed or refractory multiple myeloma. The protocol, consisting of 300 mg/day of thalidomide administered orally, intravenous incadronate (10 mg/day) administered weekly, and 12 mg/day dexamethasone for 4 days, was repeated every 3 weeks. Evaluations of efficacy and toxicity were carried out every 3 weeks and were continued for 3 cycles. Three patients were excluded during the study because of apnea, severe somnolence, and pancytopenia. Of 9 evaluated patients, the partial responses achieved in 3 patients and the minor responses achieved in 4 patients corresponded to a response rate of 78% according to the criteria of the European Group for Blood and Marrow Transplantation. In addition, painful osteolytic symptoms improved rapidly after 1 cycle of TID therapy in the 10 patients evaluated. These data suggest that TID is a feasible and promising therapeutic approach for refractory and relapsed multiple myeloma.

ADAMTS-13 activity can predict the outcome of disseminated intravascular coagulation in hematologic malignancies treated with recombinant human soluble thrombomodulin.

We conducted a multicenter prospective study for evaluating the utility and prognostic markers of recombinant human soluble thrombomodulin (rTM) treatment for acute disseminated intravascular coagulation (DIC) by various types of hematologic malignancies. The study comprised 30 patients with DIC due to hematologic diseases without severe infection. DIC improved in 15 patients and 20 were alive on day 28. Univariate analyses showed that, in comparison with patients who had survived on day 28, patients who had not survived on day 28 showed significantly higher plasma levels of plasminogen activator inhibitor‐I (PAI‐I) and significantly lower plasma activity of a disintegrin and metalloproteinase with a thrombospondin Type 1 motif, member 13 (ADAMTS‐13). Moreover, multivariate logistic regression analysis identified a significant association between plasma ADAMTS‐13 activity before treatment and survival on day 28 (P = 0.034). In particular, patients with lower ADAMTS‐13 activity (≤65%) had a poorer survival rate than those with a higher activity (P = 0.042). These findings suggest that the plasma ADAMTS‐13 activity at the time of DIC diagnosis might help to predict the prognosis of patients treated with rTM for DIC associated with hematologic malignancies. Am. J. Hematol., 2012.

Effect of vinorelbine on the growth of human myeloma cell lines in vitro

Vinorelbine (NVB) is a newly synthesized vinca alkaloid that has been used to treat advanced malignant diseases including lung adenocarcinoma and lymphoma. The effect of NVB on myeloma, however, is unknown. We therefore examined the effect of NVB on the growth of human myeloma cell lines (RPMI8226, U266 and KPMM2) using the trypan blue dye exclusion test and Alamar blue assay. NVB inhibited the growth of myeloma cells of all three cell lines dose-dependently and this effect was intensified when NVB was combined with dexamethasone at 1.0 x 10(-6)mol/l. Flow cytometric analysis using annexin V (AN) and 7-amino-actinomycin D (7AAD) showed that NVB-induced apoptosis of these myeloma cells in all the cell lines. NVB appears to be a potent inducer of apoptosis in myeloma cells, and might have some benefit in the treatment of myeloma patients.

Inflammatory pseudotumor of the spleen complicated by idiopathic thrombocytopenic purpura

Inflammatory pseudotumor is a benign disease entity of unknown etiology and pathogenesis, which mainly affects elderly patients and involves various parts of the body such as the orbit, respiratory tract, gastrointestinal tract, liver, or soft tissue. However, inflammatory pseudotumor of splenic origin has been rarely reported [1]. Here, we report a patient with splenic inflammatory pseudotumor which was simultaneously complicated by idiopathic thrombocytopenic purpura (ITP). In April 2002, a 68-year-old female was admitted to our hospital because of the bleeding tendency on her skin. She was not anemic, and neither lymph node swellings nor hepatosplenomegaly was found. Laboratory findings at that time were as follows: platelets 2×10/l, hemoglobin12.1g/dl, and white blood cells 8.1×10/l with normal differential counts. Coagulation test showed no significant abnormalities. Bone marrow was normocellular with increased number of megakaryocytes. She was serologically negative for human immunodeficiency virus (HIV). She was diagnosed as having ITP and received oral prednisolone (1.2 mg/kg). Both her platelet counts and bleeding tendency showed rapid recovery, and prednisolone was stopped in June 2002. Whereas routine ultrasonographic examination on her admission revealed the solitary hypoechoic mass (3 cm in diameter) in her spleen, no additional space-occupying lesions in the other organs or abdominal lymph node swellings were found. Concerning the splenic tumor, she had been asymptomatic and carefully observed for 10 months. However, her splenic tumor gradually enlarged and reached up to 5 cm in diameter in February 2003 (Fig. 1a). Therefore, she underwent diagnostic splenectomy. Macroscopically, her splenic mass showed a whiteyellowish cut surface with elastic firmness, which was well circumscribed but not encapsulated (Fig. 1b). Histologically, her splenic mass was composed of granulomatous nodules infiltrated by variable inflammatory cells such as lymphocytes or plasma cells. Multinuclear giant cells were also found (Fig. 1c, d). Special stains for acid-fast bacilli and fungi were negative; therefore, she was diagnosed as having splenic inflammatory pseudotumor. Epstein–Barr virus (EBV) was not detected in the tumor by in situ hybridization. She had been well without relapse of both ITP and inflammatory pseudotumor until November 2004. In the literature, most cases of splenic inflammatory pseudotumor were found incidentally, either at autopsy or during the work-up for other diseases. This is rare with only 82 reported cases in a recent literature survey [1]. Among these, there have been four reported patients (one male and three females with ages ranging from 20 to 65 years) with splenic inflammatory pseudotumor who concomitantly suffered from ITP [2, 3]. These findings suggested the autoimmune etiology of this disease. However, the clinical course of splenic inflammatory pseudotumor in our patient did not seem tobe in accordancewith that of ITP, namely, the splenic tumorhadgradually enlargedafter ITPwas alleviatedbyoral prednisolone. Alternatively, immunosuppressive therapy with prednisolone for ITPmight have triggered the progression of splenic inflammatory pseudotumor in our patient. Immunosuppressive therapy has been known to accelerate the disease progression in certain diseases such as EBVassociated post-transplant lymphoproliferative disorder. Of M. Hatsuse . S. Murakami . H. Haruyama Department of Medicine, Social Insurance Kyoto Hospital, Kyoto, Japan

Early Relapse After High-Dose Chemotherapy Rescued by Tumor-Free Autologous Peripheral Blood Stem Cells in Acute Lymphoblastic Leukemia: Importance of Monitoring for WT1-mRNA Quantitatively

A 24-year-old woman who suffered from ALL with MLL gene rearrangement received high-dose chemotherapy followed by autologous PBSC transplantation during complete remission (CR). Reverse transcriptase-polymerase chain reaction (RT-PCR) used to detect MLL/LTG4 chimeric mRNA showed no minimal residual disease (MRD) in the graft or bone marrow at the transplantation. However, the leukemia relapsed four months after transplantation. Retrospective analysis of quantitative measurement of Wilms tumor gene (WT-1) mRNA showed an increased level in the bone marrow although it was within the normal range. These observations suggest that careful monitoring of MRD by quantitative measurement of WT-1 mRNA in addition to disease-specific chimeric mRNA is required to predict relapse.

Meningeal Relapse after Double Peripheral Blood Stem Cell Transplantation in IgD Myeloma

A 54-year-old man diagnosed with IgD myeloma (stage IIIA) in complete remission (CR) received peripheral blood stem cell transplantation (PBSCT) twice with an interval of 4 months using high-dose melphalan 200   mg/m 2. However 9 months after the second PBSCT, he was readmitted because of lumbago, lower left hemiparesis, speech disturbance and left facial nerve palsy. A lumbar puncture revealed myeloma cells in the cerebrospinal fluid (CSF). The patient did not respond to any salvage chemotherapy and died of sepsis 27 months after the initial diagnosis. The findings in this patient suggest that another treatment modality including prophylactic intrathecal injection of an anti-cancer drug as well as allogeneic cell therapy is probably necessary in patients with high-risk IgD myeloma.