[Cell cycle-dependent change of the adhesive character of CD34+ progenitor cells and their VLA-4 expression].

We identified the cell cycle status of CD34+ cells of steady-state bone marrow (BM) and peripheral blood (PB) obtained from healthy volunteers, and those of BM and apheresis PB samples collected from donors who had been administered granulocyte colony-stimulating factor (G-CSF). Regardless of whether G-CSF treatment was undergone, more than 10% of CD34+ cells in the BM was in the S + G2/M phase. In contrast, less than 2% of CD34+ cells in the PB was cycling.

Differentiation in culture of murine primitive lymphohematopoietic progenitors toward T-cell lineage.

Earlier, we described a stromal cell-free two-step clonal culture system in which murine primitive lymphohematopoietic progenitors produce myeloid and B-lymphoid lineage cells. In the same culture T-cell potential of the progenitors was maintained. We now report that, in addition to myeloid and B-lymphoid cells, putative T-cell progenitors are also produced in culture.

Effects of filtration and gamma radiation on the accumulation of RANTES and transforming growth factor-beta1 in apheresis platelet concentrates during storage.

Background: Platelet-derived biologic response modifiers (BRMs) including RANTES and transforming growth factor (TGF)-beta1 accumulate in platelet components during storage because of platelet activation, and they may play a causative role in nonhemolytic febrile transfusion reactions. The majority of PCs with high unit values are provided by single donor apheresis in Japan.

Study Design And Methods: RANTES and TGF-beta1 levels in platelet units prepared from single-donor apheresis platelet concentrates (apheresis PCs) and units from whole blood (buffy coat PCs) were investigated.

A clonal culture assay for human cord blood lymphohematopoietic progenitors.

We describe a two-step clonal culture assay system for human lymphohematopoietic progenitors present in umbilical cord blood which are capable of differentiation along both myeloid and B-lymphoid lineages. Human cord blood CD34+ cells were plated in methylcellulose in the presence of stem cell factor (SCF), granulocyte-colony stimulating factor (G-CSF), interleukin (IL)-7, and the murine stroma cell line, MS-5. The growing primary colonies were individually examined for their potentials to differentiate along both myeloid and B-lymphoid lineages by reculturing aliquots of the primary colonies in methylcellulose culture containing IL-3, G-CSF and erythropoietin (Epo), and on a monolayer of MS-5 in the presence of SCF and G-CSF.

Inflammatory cytokine production in whole blood modified by liposome-encapsulated hemoglobin.

The effect of Neo Red Cells (NRC), liposome-encapsulated hemoglobin, on production of tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) were studied in whole blood preparations ex vivo. Venous blood was collected with heparin and incubated in a CO2 incubator. Treatment of blood samples with NRC reduced the constitutive levels of TNF-alpha and IL-6.

Platelet-like particle production from cultured human megakaryocytic cell line.

The human megakaryocytic cell line CMK was positive for the expression of platelet glycoprotein(GP) IIb/IIIa but few cells were positive for GPIb. To establish a GPIb-positive CMK subclone, GPIb-positive CMK cells were purified, micromanipulated and cloned, but resulting sublines were all negative for GPIb expression. The combined presence of interleukin-3 (IL-3) and the anti-virus agent ribavirin strongly induced expression of GPIb on the cells.

Effects of liposome-encapsulated hemoglobin on phorbol ester-induced superoxide production and expression of costimulatory molecules by monocytes in vitro.

The effects of Neo Red Cell (NRC), a liposome-encapsulated hemoglobin (LEH), on the phorbol ester-induced superoxide production and the expression of costimulatory molecules by human peripheral monocytes were investigated. The treatment of human mononuclear cells with NRC caused the potentiation of superoxide production in response to PMA. The longer incubation (20 h) resulted in a decrease in the PMA-induced superoxide production, which was in parallel to a decrease in the viability of the monocytes.

Lipopolysaccharide-induced desensitization of junB gene expression in a mouse macrophage-like cell line, P388D1.

Treatment of a mouse macrophage cell line, P388D1, for 1 h with bacterial LPS caused a transient increase in the level of junB mRNA expression. These cells became refractory in terms of the junB gene response to exposure to a second round of LPS or lipid A, but not to PMA. The LPS-induced desensitized state was not due to the shortening of the half-life of junB mRNA, but was suggested, by nuclear run-on analysis, to be caused by reduction of junB gene transcription.

Different adhesive characteristics and VLA-4 expression of CD34(+) progenitors in G0/G1 versus S+G2/M phases of the cell cycle.

We identified the cell cycle status of CD34(+) cells of steady-state bone marrow (BM) and peripheral blood (PB) obtained from healthy volunteers, and those of apherasis PB samples collected from healthy donors who had been administered granulocyte colony-stimulating factor (G-CSF). More than 10% of CD34(+) cells in BM were in S+G2/M phase. In contrast, regardless of whether G-CSF treatment was performed, less than 2% of CD34(+) cells in PB were cycling.

Virus inactivation in superoxide dismutase preparations by ultraviolet light irradiation.

Viral inactivation in superoxide dismutase (SOD) derived from human red cells was carried out by ultraviolet light C (UVC) irradiation. With 400 J/m2 UVC irradiation, the titer of canine parvovirus (CPV, a nonenveloped virus), M13 bacteriophage (M13, a nonenveloped phage) and vesicular stomatitis virus (VSV, an enveloped virus), which were spiked into SOD solution, were reduced by > 4.6 log10 (detection limit), 7.

Photoinactivation of virus infectivity by hypocrellin A.

We investigated the photoinactivation of virus infectivity by hypocrellin A and its mechanism. The titers of vesicular stomatitis virus (VSV) and human immunodeficiency virus type 1 (HIV-1), both of which are enveloped viruses, were reduced upon illumination with hypocrellin A in a concentration-dependent manner, whereas canine parvovirus, a nonenveloped virus, was not killed. The removal of oxygen or addition of sodium azide or beta-carotene both inhibited VSV inactivation.

[Development and clinical application of the artificial blood as the oxygen carriers].

The artificial oxygen carrier first applied in clinical treatment was Fluosol-DA, which is one of the perfluoro-chemicals. However it is not used now because of low oxygen carrying ability and immunological disturbance. On the contrary, outdated human hemoglobin is currently used by many researchers who are challenging to develop artificial oxygen carriers.

[Mistyping of ABO grouping by polyagglutination].

Polyagglutination is a phenomenon that a sample of patient's red blood cells is agglutinated by most of normal human sera. In patients with bacterial infection or hematological diseases, red cells may become agglutinable due to an exposure of antigens (cryptoantigen) that are usually hidden as submembrane structures of normal red cells. Most of normal sera contain natural antibodies to the corresponding antigens.

[Blood components: indication and notice of transfusion].

The total number of blood donors in 1996 was 6 millions, and the number of 200 ml whole blood donors was 2.16 million(35.7%), whereas the numbers of 400 ml and apheresis donors were 2.

Potential involvement of both type I and type II mechanisms in M13 virus inactivation by methylene blue photosensitization.

We have investigated the mechanism of virus photoinactivation with methylene blue (MB) by conducting deuterium oxide (D2O), azide ion (N3-) and oxygen-dependent studies. Inactivation of M13 bacteriophage and singlet oxygen (1O2) generation by MB photosensitization were irradiation dose dependent. Inactivation of M13 was enhanced by D2O and inhibited by N3-, suggesting that 1O2 participates in M13 inactivation by MB photosensitization.

Factors affecting M13 bacteriophage inactivation by methylene blue photosensitization.

We have investigated the factors that affect the virucidal activity of methylene blue (MB) photosensitization. The M13 bacteriophage was more rapidly inactivated at higher temperatures (6 degrees C < 24 degrees C < 38 degrees C). Rate constants for inactivation were 0.

Generation of interleukin 8 in stored apheresis platelet concentrates and the preventive effect of prestorage ultraviolet B radiation.

Background: Several recent studies have reported both the generation of cytokines, including interleukin (IL)-1 beta, IL-6, tumor necrosis factor alpha (TNF-alpha), and IL-8, in the supernatants of stored platelet concentrates (PCs) and the implications of this generation in febrile nonhemolytic transfusion reactions. Prestorage filtration is regarded as highly effective in the prevention of cytokine generation.

Study Design And Methods: Studies evaluated 1) the levels of these cytokines in apheresis PCs during storage, 2) the effects of white cell inactivation by ultraviolet B or gamma-radiation on the generation of cytokines, and 3) the effects of poststorage filtration on cytokine levels.

A novel gene (drad-1) expressed in hematopoiesis-supporting stromal cell lines, ST2, PA6 and A54 preadipocytes: use of mRNA differential display.

We established a differentiation-inducible preadipocyte cell line, designated A54 preadipocytes, from C3H10T1/2 (10T1/2) mouse embryo fibroblasts. A54 preadipocytes had marked hematopoiesis-supporting ability in vitro but this ability was lost after terminal differentiation to adipocytes. In this study, to identify molecules that contribute to the hematopoiesis-supporting ability of A54 preadipocytes, we screened genes that were differentially expressed in A54 preadipocytes and isolated seven novel genes by reverse transcriptase polymerase chain reaction mRNA differential display.

The impact of red cell substitutes on the blood service in Japan.

The total number of blood donors in 1994 was 6.6 millions, and 200 ml whole blood donation occupied 41.8% of all donations, whereas 400 ml and apheresis donations occupied 35.

[Basic and clinical aspects of hepatitis virus carriers].

Among the six species of hepatitis viruses, HBV (hepatitis B virus) and HCV (hepatitis C virus) can induce persistent infection. HBV and HCV are transmitted parenterally, of which maternal transmission and transfusion-associated infection is a major route respectively. We opened the special clinic for carriers detected through blood donation, and followed them at regular intervals for their health care.

Decreased inducible expression of CD80 and CD86 in human monocytes after ultraviolet-B irradiation: its involvement in inactivation of allogenecity.

Ultraviolet-B (UV-B) irradiation of antigen presenting cells (APCs) modifies their allogenecity, resulting in inhibition of the proliferative response of T cells in mixed lymphocyte reaction (MLR). Costimulation by the CD28 ligand CD80 (B7/B7-1) and CD86 (B70/B7-2) plays an important role during T-cell proliferation by augmenting synthesis of interleukin-2 (IL-2) and other cytokines. In this study, we demonstrated induced expression of both CD80 and CD86 during allogeneic MLR, though human freshly isolated monocytes express CD86 constitutively with a much lower level of CD80.

The effect of a hematopoietic-promoting factor (HPF) extracted from porcine kidney on the proliferation of human hematopoietic progenitor cells.

Hematopoietic-promoting factor (HPF), which was found in porcine kidney, has been demonstrated to act synergistically with colony-stimulating factor and erythropoietin on murine myeloid colony formation. We investigated the effect of HPF on the proliferation of human hematopoietic progenitor cells prepared from cord blood cells (CB) and peripheral blood cells (PB). HPF enhanced granulocyte colony-stimulating factor plus interleukin-3 and erythropoietin-induced colony formation, where the number of colonies were increased by 7.

Analysis of viral DNA, protein and envelope damage after methylene blue, phthalocyanine derivative or merocyanine 540 photosensitization.

Although numerous photosensitizers have been used experimentally to decontaminate viruses in cellular blood components, little is known about their mechanisms of photoinactivation. Using M13 bacteriophage and vesicular stomatitis virus (VSV) as model viruses, we have investigated alteration of the viral genome, protein and envelope after phototreatment. Methylene blue (MB) and aluminum phthalocyanine tetrasulfonate (AlPcS4) phototreatment inactivated bacteriophage M13 and decreased the fraction of single-stranded circular genomic DNA (sc-DNA) by converting it to linear form.

[Epstein-Barr virus-specific immunity in asymptomatic carriers of human T-cell leukemia virus type 1].

Adult T-cell leukemia (ATL) patients are immunosuppressed as evidenced by anergy to recall antigens and the occurrence of opportunistic infections. The immunosuppression appears to be a critical factor or a predictive sign for the development of ATL in carriers of human T-cell leukemia virus type 1 (HTLV-1). This study was aimed at assessing the immune status of asymptomatic HTLV-1 carriers with the immunity specific to Epstein-Barr virus (EBV), a ubiquitous human herpesvirus with oncogenic potential.

Human cytomegalovirus DNA is not detectable with nested double polymerase chain reaction in healthy blood donors.

The PCR method was introduced to detect cytomegalovirus (CMV) DNA from 189 peripheral blood samples of volunteer donors. We adopted the nested double PCR method with primers specific for immediate early gene 1 followed by electrophoresis and ethidium bromide staining. This nested double PCR method was sensitive enough to detect approximately a single copy of CMV DNA.