Nihon Ketsueki Gakkai zasshi : journal of Japan Haematological Society最新文献

We analyzed the phagocytic activity of purified human monocytes using fluorescent latex beads sensitized with IgG or IgG.C3 by flow cytometry. To prepare IgG-sensitized latex beads (BA), BSA-coated latex beads (B) were incubated with diluted rabbit IgG anti-BSA. To bind complement components, BA were incubated with whole serum pretreated with K-76 monocarboxylic acid (K-76COOH). K-76COOH inhibits the activity of factor I and C5, resulting in deposition of C1, C4b, C2a, C3b on BA (BAC). Phagocytic activity was assessed by percent phagocytosis and phagocytic index (PI). To eliminate the effects of non-phagocytosed latex beads, subtraction of the data at 4 degrees C from 37 degrees C was performed. Percent phagocytosis for 60 min. was B 5.0%, BA 18.3%, and BAC 57.5%, and PI (ingested latex beads/100 cells) was B 7.9, BA 36.8, and BAC 152.7, respectively. In addition, K-76COOH caused dose dependent inhibition on IgG.C3 mediated phagocytosis. Comparison of inhibition pattern on BAC and BA indicated that K-76COOH directly inhibited C3.C3-receptor binding.

We report a case of malignant histiocytosis diagnosed by liver-spleen biopsy under laparoscopy. A 49-year-old woman was admitted to our hospital with thrombocytopenia, moderate anemia and hypoproteinemia. Her bone marrow findings revealed erythroid and megakaryocyte hyperplasia, and the serum ferritin concentration was 2,250 ng/ml though she had not received any blood transfusions. Ferrokinetics analysis showed the pattern of ineffective erythropoiesis, and the half-lives of erythrocytes and platelets were both shortened. Her hepatosplenomegaly gradually increased accompanied by increasing serum ferritin level to 10,000 ng/ml. Liver-spleen biopsy was carried out under laparoscopy and revealed infiltration of atypical histiocytes with erythrophagocytosis, which were positive for S-100 and ferritin but negative for lysozyme. The rate of glycosylation in whole serum ferritin, analyzed by using concanavalin-A binding method, showed that her glycosylated ferritin content was only 8.3%, whereas in sera after iron overloading, it was about 70%. Serum isoferritin profiles by isoelectric focussing were studied, and isoferritin pattern from malignant histiocytosis was the same as that in iron overloading after neuraminidase treatment. These findings suggest that serum ferritin is synthesized in proliferating histiocytes and released in the plasma as a nonsecretory type (nonglycosylated ferritin) in this case.

A 55-year-old woman was first seen in October 1986, because of splenomegaly, moderate anemia and leukocytosis. The hemoglobin was 8.8 g/dl, platelet count 24.4 X 10(4)/microliters, and the white cell count 23,800/microliters with 73% atypical lymphoid cells. The bone marrow nucleated cell count was 99,000/microliters with 36% lymphoid cells. These atypical lymphoid cells showed hairy appearance under phase-contrast microscopy, and were positive for tartrate-resistant acid phosphatase. These cells showed the surface phenotype of CD10, CD19, CD20, Leu M5, HCM, and IgG K. Biochemical data revealed marked polyclonal hypergammagloburinemia (PHG) of IgG type (IgG 8756 mg/dl). To elucidate the mechanism of the PHG, we investigated whether hairy cells produce interleukin 6 (IL-6) and express IL-6 receptor. The culture supernatant of these hairy cells increased 3H-thymidine uptake of a IL-6 dependent hybridoma clone (MH60) in a dose-dependent manner. These cells were stained with anti-IL-6 antibody using immuno-cytochemical technique. Our results suggested that these hairy cells produce and secrete IL-6. Immunocytochemical staining with anti IL-6 receptor antibody and the binding assay with 125I-labelled recombinant IL-6 revealed that these cells express little or no receptors for IL-6. It was therefore suggested that IL-6 produced by hairy cells in this case is not an autocrine growth factor for these cells but may play a role in development of PHG by stimulating normal B lymphocytes to produce an excessive amount of immunoglobulin.

Long-term marrow cultures were established from 35 patients with aplastic anemia (AA) and the adherent stromal cell layers were assessed. Cultures from 23 of the 35 patients grew scanty stromal cell layers or did not produce any adherent cells. Long-term cultures from the remaining patients formed adherent cell layers that appeared to be morphologically normal. Cultures from 7 patients that grew apparently normal adherent cell layers were examined for expression of intermediate filament proteins using antibodies CGA-7 and HHF, which respectively recognize actin epitopes expressed in smooth muscle and normal marrow stromal cells. Cells from 4 of the 7 patients expressed vimentin (antibody 43 beta E8) but did not react with CGA-7 or HHF. It thus appears that most patients with AA have quantitative or qualitative abnormalities in the adherent cell layers from long-term marrow cultures, suggesting a defect in the hematopoietic microenvironment.

Silver staining of nucleolar organizer regions (NOR) was applied to air-dried peripheral and bone marrow smears of normal subjects and leukemic patients. Specimens were fixed in buffered acetone formalin. Even in smears kept for 2 years at room temperature, the stainability of Ag-NOR was well preserved. By dipping Giemsa-stained smears in 5% trichloracetic acid and then placing them in methanol for 5 minutes, the stain was leached out. After the dye had been removed, the smears were clearly stained by a Ag-NOR staining technique. The mean number of Ag-NOR per nucleus of mature granulocytes and mononuclear cells in normal peripheral bloods was 0.59 and 1.43 respectively. The mean number of Ag-NOR per nucleus of peripheral and bone marrow leukemic cells from patients with acute leukemia and chronic myelocytic leukemia in blastic crisis was 2.32 and 2.66 respectively. On the other hand, the mean number of Ag-NOR per nucleus of peripheral leukemic cells from patients with chronic lymphocytic leukemia was 1.48. These results suggest that acute leukemia cells possess a more active proliferating potential. The Ag-NOR staining technique is very simple and might be useful for investigation of hematologic cells.

We conducted a study on autoinduction of differentiation in human myelocytic leukemia cells (HL-60-Y3) in which the effects of serum cytodifferentiation were excluded by the use of a serum-free semisolid culture. In the culture dish the HL-60-Y3 colony count per dish was kept at 100 or below, and only the formation of clumping-type colonies, which consisted of blastoid cells, was observed. The formation of spreading-type colonies increased with the colony count and when the colony count reached 500 per dish, more than 90% of the colonies formed were spreading-type colonies. The main component cells of the spreading-type colonies were mature monocytoid cells, which were positive for alpha-naphthyl butyrate esterase. Moreover, a marked reduction in the recloning ability was observed in differentiated colonies compared to undifferentiated colonies. These results indicate the autoinduction of differentiation in human myelocytic leukemia cells. Furthermore, a single cell study that excluded the effect of colony to colony interactions suggested the presence of a differentiation autoinducing factor in the medium.

beta-Glucuronidase purified from normal human tissues (placenta, liver and spleen) and granulocytes was composed of 80 kilo-Dalton (kDa), 64 kDa and 18 kDa subunits. The enzyme from human myelogenous leukemic cells contained a 80 kDa subunit as a major component. Amino acid sequencing revealed that N-terminal regions of the 80 kDa from CML cells and the 18 kDa from placenta were identical, and the sequence of the 64 kDa from placenta was identical to the downstream sequence of Gly138 in the 80 kDa enzyme. Therefore, it is probable that the 80 kDa is a precursor form and the 64 kDa is a mature form that is derived by removal of N-terminal 18 kDa peptide (137 amino acids) from the precursor form. These observation simply that proteolytic processing of this enzyme is impaired in myelogenous leukemic cells. The possibility of impaired protease activity against the 80 kDa subunit in CML cells was excluded, since the 80 kDa was not susceptible to several protease sources from normal tissues (placenta and liver) and granulocytes under a variety of reaction conditions. In addition no mutation of the primary structure in myelogenous leukemic cells was detected through analysis of DNA encoding for a peptide, including Gly138 of the enzyme. Therefore, it is suggested that abnormal processing of beta-GUase in the myelogenous leukemic cells is due to alteration in transcriptional or translational step.

Thirty-eight patients with chronic refractory idiopathic thrombocytopenic purpura (ITP) were treated with weekly slow infusions of vincristine (0.02 to 0.04 mg/kg) or vinblastine (0.1 to 0.2 mg/kg). Twenty-two patients showed good to excellent responses after one to eight infusions. These responses were generally short, and lasted only in six patients after discontinuance of the therapy. The efficacy was comparable between vincristine and vinblastine. Neither the age, sex, duration of the disease, prior splenectomy nor combined use of adrenocortical steroids was likely to have influenced the therapeutic effect. Side effects such as peripheral neuropathy, alopecia, gastrointestinal symptoms and leukopenia occurred in 34 patients, and necessitated discontinuance of the therapy in eight patients. Slow infusions of vinca alkaloids can be an effective means of inducing platelet response in patients with chronic refractory ITP, but frequent side effects limit its clinical usefulness.