The neutrophilic leukocyte has proved to be a valuable model for studying signal transduction pathways. Neutrophil responses proceed from recognition of appropriate stimuli to an activation phase that initiates the terminal activation sequences involving activated NADPH oxidase, chemotaxis, and enzyme secretion. Oxidase activation in neutrophils involves the assembly, in the plasma membrane, of membrane-bound and cytosolic components of the oxidase complex, which were disassembled in their resting state. Chemotaxis involves actin, microtubles and plasma protein (ex. gelsolin). In these processes, tyrosine phosphorylation, G-protein and activation of phospholipases play important roles. The present review is focused on recent data concerning the signaling pathways which lead to oxidase activation and cellular motility, including pathogenesis and the biochemical aspects of diseases.

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The major source of transplantable hematopoietic stem cells (SC) are presently the bone marrow (BM) and peripheral blood after intensive chemotherapy. Recently, human cord blood (CB) as a source of transplantable SC has shed light on SC transplantation. We studied the total number of collectable SC from each of 28 placenta. CB were harvested from umbilical cord veins of placentae immediately after normal deliveries. Red blood cells of CB were removed by the addition of hydroxyethyl starch, and then supernatant containing leukocytes and SC were collected. The number of SC in the collected CB were evaluated by an in vitro colony assay. The total number of CFU-GM of CB ranged from 0.8 to 33.1×10⁵ (median 4.5×10⁵), BFU-E were from 1.1-35.1×10⁵ (median 4.1×10⁵), and CFU-mix were from 0.15-7.23×10⁵ (median 0.57×10⁵). Although variability was noted between CB samples, it was clear that CB contained substantial numbers of SC. The results suggested that CB harvested from placentae contained a large number of SC and that there is probably a sufficient number of SC in a single CB collection to reconstitute the BM of children.

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The bone marrow microenvironment components following allogeneic bone marrow transplantation (BMT) are reported to be of the recipient origin, implying that the marrow components can not be transplantable in vivo. To investigate the transplantability of the marrow microenvironment components, we have constructed an in vitro model of transplantation using 12 pairs of long-term cultured fibroblastic stromal cells obtained from different individuals. In our in vitro model of transplantation, after ablation of all hemopoietic cells, we simultaneously established a pair of confluent layers of fibroblastic stromal cells from different individuals and transplanted one of them on the other confluent layer cells. The overlaid stromal cells grow on the allogeneic confluent layer as observed by phase-contrast microscopy. The immunostaining showed that the stromal cells were vimentin-positive fibroblasts. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) showed mixed chimerism in all 12 pairs. The chimerism was found if the dose of overlaid stromal cells was more than one-tenth that of the confluent layer cells. These data indicate that fibroblastic stromal cells are transplantable in vitro even if the surface of the flask is completely covered with the allogeneic stromal cells, and their dose may be largely related to their transplantability following allogeneic BMT in humans.

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We compared bone marrow suppression and the incidence of infection between patients who received pyrarubicin (THP) concomitantly with vincristine, prednisolone and L-asparaginase and patients who received adriamycin (ADM) concomitantly with vincristine, prednisolone and L-asparaginase for the induction of remission in childhood acute lymphoblastic leukemia. In the ADM group, the duration of neutropenia lower than 500/μ1 and lower than 200/μ1 was 13.7±9.9 days and 8.4±7.3 days, respectively. The duration of such neutropenia in the THP group, 23.9±5.1 days and 20.0±8.9 days, respectively, was significantly longer than that in the ADM group. In the ADM group, infection occurred in one of 12 patients, whereas, in the THP group, infection occurred in seven of 9 patients, and one patient died. The protocols had to be modified or interrupted in these seven patients. The incidence of infection in the two groups was significantly different. THP caused greater bone marrow suppression than ADM. Thus, we recommend strict prophylaxis for infections in patients who are to receive THP.

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We describe eight children with hemophagocytic syndrome (HPS) : malignant histiocytosis, 1 case ; familial erythrophagocytic lymphohistiocytosis (FEL), 1 case ; virus-associated HPS, 3 cases (Epstein-Barr virus, 1 case and cytomegalovirus, 2 cases); infection-associated HPS, 1 case (mycoplasma); each 1 case of Still's disease and hybrid leukemia with HPS. Three cases in which bone marrow proliferation of immature atypical histiocytes could be detected, demonstrated severe pancytopenia and organ failure with a prominent hypercytokinemia (IL-6, IFN-γ, sIL-2R). Six patients are alive with no evidence of disease from 12 to 57 months (median, 27 months) after the time of diagnosis. This study indicates that the prognostic prediction should be done by taking into account both bone marrow morphology (Manoharan's classification) and the staging system (the Imashuku's or Oka's).

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A 9-year-old girl with glioblastoma multiforme was treated with surgical resection (95%) and high-dose multiple drug chemotherapy followed by autologous bone marrow transplantation. Nimustine, etoposide, vincristine, methotrexate and carboplatin were administrated by intravenous injection and procarbazine was administrated orally for 7 days. The nucleated marrow cells, 0.85 × 10⁸ cells/kg (0.32×10⁵ CFU-GM/kg), were infused. The patient recieved granulocyte colony-stimulating factor from day 1 to day 22. The absolute neutrophil count reached 500/μ1 on day 13 and the platelet count reached 50, 000/μ1 on day 28. Red blood cell transfusion and platelet transfusion were performed on days 3 and 4, respectively. Febrile episodes were observed during periods of neutropenia for 18 days. Reversible toxicities to heart and kidney were observed.She has been in complete remission for 3 years and 1 month. We suggest this treatment method is useful for patients with GBM.

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A 6-month-old male infant was referred for evaluation of anemia. Severe iron deficiency anemia, respiratory distress with increased lung field density dominantly in dorsal zone of chest CT images, and hemosiderin-laden macrophage (siderophage) in the gastric juice confirmed a diagnosis of IPH. In the management of our patient, the significant observations were : (1) the efficacy of corticosteroid “pulse” therapy for reducing acute exacerbation of IPH; (2) the possibility of intravenous liposteroid (corticosteroid incorporated in lipid emulsion) to prevent relapse. Liposteroid has a specific drug-delivery system so that corticosteroid can act mainly in inflammatory tissue. We consider that liposteroid may provide good long-term control of the disease.

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A 10-year-old girl with aplastic anemia, was complicated with pure red cell aplasia (PRCA) after major ABO mismatched allogeneic bone marrow transplantation is reported herein. She received major ABO mismatched bone marrow (donor A, Rh+ ; recipient O, Rh+) from her mother whose HLA was phenotypically identical to the recipient. Her post-transplant course was uneventful except for the lack of erythroid recovery. She was diagnosed as PRCA because of marked erythroid hypoplasia of bone marrow cells. Nine months after BMT, high-dose erythropoietin was administered subcutaneously, which did not show any effect on proliferating erythroid cells. Fourteen months after BMT, we tried high-dose gamma-globulin (IVIG) therapy (400 mg/kg, 5 consecutive days). Soon after the IVIG therapy, her reticulocyte count elevated markedly. The myelogram on day 4 after the IVIG therapy showed marked erythroid hyperplasia. She achieved durable remission thereafter.

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A 14-year-old boy with acute lymphoblastic leukemia with t (4;11) (q21;q23), relapsed in the marrow while on therapy after 8 months of complete remission, and received double syngeneic transplantation. First, for the purpose of induction, he received preconditioning with BU+VP-16+ CY and transplanted a half of the harvested marrow cells ; the other half was cryopreserved for the second transplant. Thirty days after the first transplant, he entered remission and on 65 days restarted maintenance therapy (PSL+VCR+6-MP+MTX).Six months after the first transplant, he received preconditioning with TBI+L-PAM and the second transplantation. The cryopreseved marrow and peripheral blood mononuclear cells collected after the administration of recombinant human granulocyte colony-stimulating factor (G-CSF) to the donor were transfused together for earlier blood cell recovery and escaping from complications. Currently, he has been surviving disease-free for 10 months without maintenance therapy. In conclusion, double transplantation may be an effective strategy for refractory poor prognosis leukemia, and transplantation with peripheral blood mononuclear cells collected after the administration of G-CSF to the donor may be useful.

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We report the case of a 1 year and 4 month old girl who needed repeated red blood cell transfusions during 6 months after autologous bone marrow transplantation (ABMT) for neuroblastoma (stage IV A). Original tumors were removed after 3 cycles of A 1 protocol, and additional systemic chemotherapies were done. The conditioning therapy was high-dose chemotherapy and total body irradiation. The time reaching absolute granulocyte counts over 500/μl and white blood cell counts over 1, 000/μl after ABMT was 20 days and 21 days, respectively. Because platelet and red blood cell recoveries were delayed, the patient needed repeated platelet and red blood cell transfusions. Treatment with M-CSF was started 107 days after ABMT. Seven days after treatment with M-CSF, edema was noted. Treatment with M-CSF was stopped, and hypertension, hematuria, proteinuria developed thereafter. After recovery of renal dysfunction, severe anemia continued and required repeated red blood cell transfusions. Endogenous erythropoietin response to anemia was inadequately low because of no response of erythropoietin production to lower hematocrit levels, and administration of exogenous recombinant human erythropoietin was effective in correcting anemia.

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