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

PHA-leukocytes conditioned medium (PHA-LCM) prepared from the patients with chronic renal failure and normal control was tested for BPA by progenitor cell assay with methylcellulose culture method. The BPA of uremic patients, expressed as a percentage of standard CM, was significantly lower than that of normal subjects (96 +/- 9%). However, there was no correlation between BPA and the severity of anemia. The number of circulating BFU-E per milliliter of blood was significantly lower in uremic patients (71 +/- 77) than in normal controls (131 +/- 96), and the number correlated with the severity of anemia. From these results, the maturation process of erythroid series in uremic patients appeared to be impaired at a stage between pluripotent stem cell and BFU-E, and might be secondary to inefficient production of BPA by lymphocytes.

Sera from 5,705 persons were tested for antibodies to human T-cell leukemia virus type 1 (HTLV-I) by gelatin particle agglutination test (PA) and enzyme immunoassay (EIA) as screening methods, and the antibodies were confirmed by indirect immunofluorescence assay (IF) and Western blotting method (WB). The antibodies were detected in 30 (1.34%) of 2,239 patients who had received blood transfusions, 3 (0.29%) of 1,022 healthy blood donors, all 18 adult T-cell leukemia/lymphoma (ATL/L) patients, 23 (47%) of 49 family members of the ATL/L patients and 10 (0.53%) of the other 1,898 persons. Before the introduction of the mass screening of healthy blood donors by the Japan Red Cross Blood Center, 17 (2.29%) of 743 recipients tested were positive for the antibodies by PA and most of them were confirmed by IF and WB. Ten of these 17 recipients tested were negative for the antibodies before the blood transfusions. After the introduction of mass screening, 13 (0.87%) of 1,496 recipients tested were positive for the antibodies by PA, but the seroconversion of the antibodies by blood transfusions could not be confirmed clearly. In persons with low titers (1:16-1:64) in PA, the coincident rate of positive reaction with EIA, IF and WB was only 9.5%, 0.0% and 23.8%, respectively, and most of the immunoglobulin (Ig) class was IgM. On the other hand, in those with titers of 1:128 or greater in PA, the coincident rate was 72.7%, 81.8% and 90.9%, respectively, and the Ig class was IgG or IgG + IgM.

Spontaneous complete remission of four months' duration was observed in a 51-year-old male with hypoplastic leukemia. Cytogenetical analysis revealed that leukemia cells of this patient were tetraploid. The diameter of leukemia cells involving myeloid cells ranged from 30 to 50 mu. The remission was apparently associated with repeated blood transfusions and severe infection. Complete remission was confirmed by normal morphology and karyotype of the bone marrow cells, although in vitro marrow stem cell growth did not return to normal. Thus, normal hematopoiesis may not have recovered when the diagnosis of spontaneous remission was made.

We used flow cytometry to investigate the change of platelet membrane glycoproteins (GPIb and GP IIb/IIIa) and the distributions of fibrinogen (Fbg), thrombospondin (TSP) and fibronectin (Fn) on the surface of thrombin-stimulated platelets. The binding of a monoclonal antibody directed at the von Willebrand factor binding site on GPIb decreased in thrombin-stimulated platelets. This antibody caused a reactive delay in thrombin-induced aggregation, but had little influence on aggregability. Slight thrombin-induced aggregation was observed even after blocking the binding of Fbg to GP II b/IIIa. The new expression of GP II b/IIIa was detected on the surface of thrombin-stimulated platelets, whereas there was little increase of Fbg dependent on this GP II b/IIIa. An increase of TSP after thrombin stimulation was observed on the surface of platelets of healthy controls and patients with Glanzmann's thrombasthenia (Type I). The level of on platelet surface was slightly increased by thrombin stimulation. The mechanism involved in thrombin-induced aggregation appears to differ from that in ADP-induced aggregation.

Mechanisms of aggregation and release reactions of human platelets induced by non-hemolytic influenza virus were studied. The influenza virus (PR/8 strain) caused aggregation of platelets with ATP release in a dose-dependent manner over the range of 640-10,240 HA titers. The aggregation was always preceded by a lag period and subsequent shape changes. The virus-induced aggregation was enhanced by exposure of the reaction mixtures to cold at 4 degrees C for 30 min and inhibited by apyrase, acetylsalicylic acid and dipyridamole. Ingestion of acetylsalicylic acid also inhibited the aggregation. Gel-filtered platelets were aggregated by influenza virus only after the reaction mixtures had been previously incubated at 4 degrees C for 30 min and then stirred at 37 degrees C. In the absence of divalent cations (Ca2+ less than or equal to 2 x 10(-5) M, Mg2+ less than or equal to 1 x 10(-5) M), gel-filtered platelets were not aggregated by influenza virus. These results suggest that influenza virus was absorbed onto the platelet surface and caused the release of ADP from platelets, which in turn, aggregated platelets.

We investigated the location of platelet-specific alloantigen Baka on platelet membrane glycoproteins. In indirect immunoprecipitation experiments, the anti-Baka antibody precipitated glycoprotein (GP) II b and a small amount of GP III a. The immunoblots using partially purified GP II b/III a complex as the target antigen indicated that GP II b alpha carried the Baka alloantigen. When the partially purified GP II b/III a complex digested with chymotrypsin was examined, the Baka alloantigen was found on a 65 kD fragment derived from GP II b alpha under reducing conditions. In addition, the immunoblots after two-dimensional nonreduced-reduced SDS-PAGE directly indicated that the 65 kD fragment had a mol. wt. of 80 kD under nonreducing conditions. The immunoblots using platelets digested in situ with chymotrypsin indicated that the 65 kD fragment of GP II b alpha was retained by the platelet membrane. We conclude, therefore, that the Baka alloantigen is located on a 65 kD fragment that represents the membrane side of the cleavage site of chymotrypsin on GP II b alpha.

We tested purified recombinant hemopoietic factors for their effects on the proliferation and differentiation of murine myelomonocytic leukemia cells (WEHI-3B-Y1) in serum-free agar culture. We found that purified recombinant human G colony-stimulating factor (CSF) markedly increased the colony number of WEHI-3B-Y1 cells and differentiation-inducing activity. However, at low colony densities [( 100/dish), G-CSF did not induce the differentiation of WEHI-3B-Y1 cells. We conclude that G-CSF does not induce the differentiation of WEHI-3B-Y1 cells directly, but induce the differentiation as a result of the secondary autoinduction of differentiation. Interleukin 3 (IL-3) slightly enhanced but erythropoietin (Epo) did not alter the colony number of WEHI-3B-Y1 cells. GM-CSF or M-CSF decreased the colony number of WEHI-3B-Y1 cells. Such purified recombinant human IL-3, Epo, GM-CSF and M-CSF did not induce morphologically the distinct differentiation of WEHI-3B-Y1 cells.

Alloreactive mixed lymphocyte reaction (MLR), mitogen-induced response (MR), and suppressor cells against these responses in murine bone marrow chimeras were examined, to clarify the mechanisms of immunological tolerance and immunodeficiency after bone marrow transplantation (BMT). Between 35 and 70 days after BMT, there was no response of spleen cells from allogeneic chimeras against host (C3H/He) and donor (BALB/c) cells, although responses against third party (C57BL/6) cells were detected, thus indicating that these allogeneic chimeras were immunologically tolerant. The activity of suppressor cells against alloreactive responses was increased 35 to 55 days after BMT, so that these suppressor cells appeared to be related to immunological tolerance. Some of the suppressor cells against alloreactive responses were Thy1+. Among them some were Lyt1+ or Lyt2+, and others were Lyt1+2+. Plastic dish non-adherent cells had slightly weaker suppressor activity than adherent cells. Proliferative responses to Con A, PHA, and PWM were decreased 13 to 15 days after BMT, and gradually increased. The responses to LPS differed from those to the former three mitogens, showing an enhanced response 21 to 25 days after BMT. The increased response to LPS did not appear to be simply due to the increased number of non-T cells in the spleen of allogeneic chimeras. The alloantigen specific suppressor cells may play an important role in the induction and maintenance of immunological tolerance, while the alloantigen nonspecific suppressor cells and suppressor cells against MRs may be related to immunodeficiency after BMT.

Cystine was transported into human erythrocytes in the presence of tertiary-butyl hydroperoxide (t-BH) or 1-chloro-2, 4-dinitrobenzene (CDNB). The transport rate of cystine was dependent on the extracellular concentration of t-BH or CDNB, and on the incubation time. By Dowex-1 column chromatography, the transported cystine was incorporated into fractions of glutathione disulfide (GSSG) and glutathione-S (GSH-S) conjugate. Cystine was also transported into reconstituted erythrocyte ghost with GSSG. The transport of cystine was Na+ dependent and decreased in the presence of N-ethylmaleimide, and it was competitively inhibited by DL-homocystine and L-alanine. The inhibition rates by DL-homocystine and L-alanine were 75% and 68%, with similar Ki values of 0.7 mM and 0.6 mM, respectively. The Km value for cystine transport was 0.15 mM. The activity of GSH-cystine transhydrogenase was detected in the hemolysate and this enzyme is thought to catalyze the action of incorporation of cystine into GSH. This enzyme was partially purified from normal human erythrocytes. In the presence of CDNB, similar rates of cystine transport were observed among the diabetic patients (n = 11), hypoxemic patients (n = 10) and the control subjects (n = 20). It is suggested that cystine transport is induced for glutathione synthesis when human erythrocytes are exposed to oxidative stresses.

The intracytoplasmic localization of CD3 antigen was examined in azurophilic granular T-lymphoblastic lymphoma cells obtained from a 27-year-old female. Examination by flow cytometry and an immunohistological method revealed that the surface phenotype of the lymphoma cells were NKH1+, CD5+, CD7+, CD1-, CD2-, CD4-, CD8- and CD16-. CD3 was not detected on the cell-surface by flow cytometry but was found in the cytoplasm using the immunohistological method. CD3 was detected in the perinuclear space and rough endoplasmic reticulum (rough ER) by an immunoelectron microscopic study. An electron microscopic observation of the granules revealed lysosomal structures with low electron density. The lymphoma cells proliferated when cultured in the presence of interleukin-2 but showed no NK (K562) activity before and after culture. The molecular size of metabolically labeled intracytoplasmic CD3 precipitated by Leu 4 monoclonal antibody was identical to that of normal CD3. These results indicate that the lymphoma cells are neoplastic counterparts of immature thymocyte at a very early stage of differentiation and may be related to the lineage of CD3+ NKH1+ T-lymphocyte.