bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of NK-lineage markers on peripheral blood lymphocytes with T- helper (Leu3+/T4+) phenotype in B cell chronic lymphocytic leukemia
Authors: Velardi A , Prchal JT , Prasthofer EF , Grossi CE 
Published: October 2019

Abstract
Heterogeneity within lymphocyte subsets expressing T-helper (T4+/Leu3+) or T-suppressor (T8+/Leu2+) markers was analyzed in 38 patients with B cell chronic lymphocytic leukemia (B-CLL) and in 11 age-matched controls. Co-expression of NK-lineage markers (M1, Leu7) on Leu2+ or Leu3+ cells was investigated by two-color immunofluorescence, and the proportion of granular lymphocytes within each subset was determined by cytochemical staining for acid phosphatase. B-CLL patients and normal controls had similar absolute numbers of cells per microL with T- suppressor phenotype. However, the proportion of Leu2+ cells co- expressing the Leu7 antigen was higher in the B-CLL patients than in the control subjects (54 +/- 3% v 27 +/- 4%, P less than .0001). The absolute number per microL of cells with T-helper phenotype was somewhat decreased in B-CLL patients compared with normal subjects (649 +/- 104 v 799 +/- 33, P less than .02), with a consequent decrease of the helper/suppressor ratio. Furthermore, co-expression of the Leu7 and, more strikingly, of the M1 markers was increased significantly on Leu3+ cells from B-CLL patients compared with normal controls (11 +/- 2% v 2 +/- 0.7%, P less than .002 for Leu7 and 40 +/- 5% v 4 +/- 1%, P less than .00001 for M1). Cytochemical studies showed that a large proportion of Leu3+ cells from B-CLL patients were granular lymphocytes, as suggested by the co-expression of natural killer (NK) cell markers. The emergence of a population of Leu3+ granular lymphocytes with NK markers, which is barely detectable in normal subjects, may provide an explanation for the impairment of T cell functions repeatedly described in B-CLL.
Lithium stimulation of granulopoiesis in diffusion chambers--a model of a humoral, indirect stimulation of stem cell proliferation
Authors: Doukas MA , Niskanen EO , Quesenberry PJ 
Published: October 2019

Abstract
Lithium has been recognized as a stimulator of granulopoiesis both in vivo and in vitro. The mechanism by which lithium provokes this stimulation is unclear, with previous data focusing on such divergent causes as direct effects on progenitor cells v elevations in granulocyte macrophage colony-stimulating activity (GM-CSA) production. In the present study, we used a model system of granulopoiesis in diffusion chambers to study this stimulation of granulopoiesis. Lithium pretreatment of mice followed by a rest period to allow for excretion of the lithium (confirmed by serum assays) revealed a stimulation of progenitor cell growth within the diffusion chambers. No changes in the serum and chamber fluid GM-CSA levels were discernible between the control host mice and the lithium-pretreated mice. These data indicate that lithium stimulates granulopoiesis by an indirect mechanism that does not appear to involve GM-CSA.
Macrophage factor X activator formation: metabolic requirements for synthesis of components
Authors: Shands JW Jr 
Published: October 2019

Abstract
The in vitro production of factor VII-like material and of tissue factor activity by murine thioglycollate exudate macrophages was measured by amidolytic assays. Tissue factor activity was inducible by endotoxin, and its induction was inhibited by 1 microgram/mL of actinomycin D, 10 micrograms/mL of cycloheximide, and 0.2 micrograms/mL of tunicamycin. Soluble factor VII-like material was secreted by macrophages into culture supernatants. The amount produced was not influenced by further activation of the cells by endotoxin, nor was its production inhibited significantly by 1 microgram/mL actinomycin D or 0.2 micrograms/mL tunicamycin. The production of the factor VII-like material was inhibited by 10 micrograms/mL of cycloheximide, and its appearance in culture supernatants was enhanced significantly by the addition of vitamin K1. When lysates of activated macrophages were suspended in ultracentrifuged culture supernatants, a particulate factor X activator was formed. Centrifugation at 100,000 g pelleted the factor X activator and left no factor VII-like material in the supernatant. The data indicate that thioglycollate-induced exudate macrophages make and excrete factor VII-like material, and this production is not modulated by further activation. However, activation of the macrophages induces tissue factor production. The factor X activator appears to result from the interaction and complexing of the soluble factor VII-like material and the membrane-bound tissue factor.
The relative importance of thrombin inhibition and factor Xa inhibition to the antithrombotic effects of heparin
Authors: Buchanan MR , Boneu B , Ofosu F , Hirsh J 
Published: October 2019

Abstract
The relative importance of antithrombin and anti-factor Xa activities of heparin fractions required to achieve optimal antithrombotic effects is unknown. To study this, we measured the effects of standard heparin, an octasaccharide heparin fraction (anti-factor Xa activity only), and dermatan sulfate (antithrombin activity only) on the prevention of thrombosis and related this to their anticoagulant effects in vivo in rabbits. Thrombosis was measured as the incorporation of 125I- fibrinogen into tissue thromboplastin-induced thrombi using a Wessler- type model. Ex vivo changes in thrombin clotting time (TCT) were used as an index of antithrombin activity, and a chromogenic anti-factor Xa assay was used to measure anti-factor Xa activity. In addition, the ability of the three sulfated polysaccharides to simultaneously inhibit the generation of thrombin activity and to enhance the inactivation of the factor Xa added to initiate thrombin generation in plasma was determined. Standard heparin, in a dose of 10 anti-factor Xa U/kg, inhibited thrombus formation by 90%, prolonged the TCT by two seconds, and resulted in an anti-factor Xa level of 0.32 U/mL. The octasaccharide heparin fraction, in a dose of 10 anti-factor Xa U/kg, inhibited thrombus formation by 41%, had no effect on the TCT, and resulted in an anti-factor Xa level of 0.28 U/mL. Higher doses of the octasaccharide resulted in a further increase in the anti-factor Xa levels but had no further effect on thrombus formation. Dermatan sulfate, in a dose of 500 micrograms/kg, inhibited thrombus formation by 95%, but had no affect on the TCT. These results indicate that the antithrombotic effect achieved by inhibiting factor Xa is limited and that better antithrombotic effects are achieved by heparin or heparin- like substances capable of influencing the inactivation and/or the generation of thrombin.
Poor response to danazol in hemophilia
Authors: Kasper CK , Boylen AL 
Published: October 2019

Abstract
We gave danazol (600 mg/day orally for 14 days) to eight adults with mild or moderate hemophilia A, one with severe hemophilia A, and one with moderate hemophilia B. In the patient with severe hemophilia A, the levels of factor VIII two to four days after an infusion of factor VIII concentrate were higher than expected, suggesting a prolonged half- life. In one patient with mild hemophilia A, a questionable slight increase in factor VIII was noted at the end of the study. No change was seen in factor levels of other subjects. Therapy was terminated early, at eight days, in a patient who developed severe muscle cramps, and at ten days in a patient with a severe rash. Another patient developed hepatic dysfunction three days after completing the 14-day trial. In this trial, the side effects of danazol outweighed its meager and questionable benefits.
Volume-dependent human blood polymorphonuclear leukocyte heterogeneity demonstrated with counterflow centrifugal elutriation
Authors: Berkow RL , Baehner RL 
Published: October 2019

Abstract
Human peripheral blood polymorphonuclear leukocytes (PMNs) have recently been recognized as a heterogeneous population of cells. Consideration has not been given to the possibility that size may be an additional physical characteristic demonstrating heterogeneity. Using counterflow centrifugal elutriation, we have demonstrated that PMNs can be isolated into at least six volume-dependent fractions. A positive correlation exists for PMN size and superoxide anion release upon stimulation with f-Met-Leu-Phe or phorbol myristate acetate. Total granule contents were also noted to be greater in larger PMN fractions, with a constant percent of release upon stimulation. The implications of these findings are discussed.
Multi-CSF-dependent colony formation by cells of a murine hemopoietic cell line: specificity and action of multi-CSF
Authors: Metcalf D 
Published: October 2019

Abstract
Cells of the Multi-CSF (IL-3)-dependent hemopoietic cell line 32D c13 formed colonies of varying size in agar cultures stimulated by Multi- CSF. Colony formation was linear with respect to cultured cell numbers; colony numbers and size increased with increasing concentrations of Multi-CSF, and 32D colonies themselves contained a high frequency of clonogenic cells. Clonogenic 32D cells died in the absence of Multi-CSF (half-life six hours), and most were unable to complete cell cycles in progress at the time of withdrawal of Multi-CSF. The concentration of Multi-CSF directly influenced the length of the cell cycle of dividing 32D cells. Purified GM-CSF, G-CSF, or M-CSF had no capacity to support the survival or proliferation of 32D cells. Colonies formed by 32D cells appear to offer a useful model for analyzing the action of Multi- CSF in controlling self-renewal by clonogenic hemopoietic cells.
Regulation of human peripheral blood erythroid burst-forming unit growth by T lymphocytes and T lymphocyte subpopulations defined by OKT4 and OKT8 monoclonal antibodies
Authors: Wisniewski D , Strife A , Wachter M , Clarkson B 
Published: October 2019

Abstract
To reexamine the influence that T lymphocytes have on the regulation of human peripheral blood burst-forming unit (BFU-E) proliferation in the absence of a statistically significant number of monocytes, very low numbers (3 to 10 X 10(3)/mL) of a null cell fraction highly enriched for BFU-E were cultured alone and in the presence of 5 X 10(5) sheep erythrocyte-purified, autologous T lymphocytes in a methylcellulose culture system containing erythropoietin. T lymphocytes consistently enhanced the growth of BFU-E from the null cell fraction, as reflected in both their number and size. Irradiation of T lymphocytes prior to coculture with null cells markedly reduced this enhancement, strongly suggesting that T lymphocytes synthesize erythroid burst-promoting factors (BPA). To determine whether there were functional differences between the two major T lymphocyte populations as defined by OKT4 (T helper/inducer) and OKT8 (T suppressor/cytotoxic) murine monoclonal antibodies to stimulate the growth of BFU-E, both T cell subpopulations were isolated by negative (panning) or positive (fluorescence-activated cell sorting) selection and cocultured with null cells. No statistically significant differences emerged between unseparated, OKT4+ and OKT8+ T lymphocytes in their ability to stimulate the growth of BFU-E. Thus, these studies provide further evidence that T lymphocytes are a major population of BPA-producing cells and further that OKT4+ and OKT8+ T lymphocytes equally elaborate these factors.
Fibrinogen biosynthesis in isolated guinea pig megakaryocytes
Authors: Leven RM , Schick PK , Budzynski AZ 
Published: October 2019

Abstract
Fibrinogen synthesis was investigated in guinea pig megakaryocytes. Purified megakaryocytes were incubated with 35S-methionine in methionine-free incubation medium for 18 hours. Newly synthesized fibrinogen in megakaryocyte lysates enriched with purified carrier guinea pig fibrinogen was immunoprecipitated with a specific anti- guinea pig fibrinogen antiserum produced in rabbits. Proteins in the immunoprecipitates were analyzed with a 3.5% to 10.0% gradient polyacrylamide slab gel electrophoresis and auto-radiography. Radioactivity was detected in a protein band of 340,000 daltons. In order to verify fibrinogen synthesis, immunoprecipitate was analyzed by two-dimensional slab gel electrophoresis: (1) the first dimension separated unreduced fibrinogen using a 3.5% to 10.0% gradient gel; (2) following reduction by 2-beta-mercaptoethanol, fibrinogen chains were separated in the second dimension using a 10% gel. Alpha, beta, and gamma fibrinogen chains, which represented carrier guinea pig plasma fibrinogen, were visualized by Coomassie brilliant blue. Autoradiography identified the incorporation of radioactivity into the three fibrinogen chains. In control experiments, immunoprecipitates, produced by exposing megakaryocyte lysates to preimmune rabbit serum and goat anti-rabbit IgG, were also analyzed by the two-dimensional gel system. Radioactivity was not detected in sites corresponding to the migration of fibrinogen subunits. The study demonstrates that isolated guinea pig megakaryocytes can synthesize fibrinogen. The electrophoretic mobility of newly synthesized fibrinogen and subunits is similar to that of guinea pig plasma fibrinogen.
Down-regulation of K cell activity by neutrophils
Authors: Dallegri F , Patrone F , Frumento G , Ballestrero A , Sacchetti C 
Published: October 2019

Abstract
Human neutrophils, activated by phorbol-myristate acetate (PMA), (A- neutrophils), were found to suppress lymphocytic killer (K) cell- mediated antibody-dependent cellular cytotoxicity (ADCC). Resting (R) neutrophils, ie, PMA-untreated cells, were completely ineffective. Suppression was optimal when A-neutrophils were added at the beginning of the ADCC assay. Furthermore, A-neutrophils were found to cause an approximately 80% reduction in the number of Raji target cell-bound lymphocytes. These data indicate that A-neutrophils inhibit K cell activity by interfering with the target cell recognition. A-neutrophils were capable of reducing the percentage of Fc receptor (FcR)-bearing lymphocytes with a half-time of 7.2 minutes, through a process preventable by the serine-protease inhibitors tosyl-lysine-chloromethyl ketone (TLCK) and lima bean trypsin inhibitor (LBTI). Conversely, A- neutrophils caused a very slow decrease in the amount of Raji cell- bound antibodies, as detected by the complement-mediated lytic assay. Thus, only lymphocyte FcR structures seem to be highly susceptible to neutrophil-derived TLCK- and LBTI-inhibitable proteases. Furthermore, supernatants from A-neutrophils were found to inhibit K cell ADCC and lymphocyte binding to Raji target cells. In addition, LBTI prevented the A-neutrophil-dependent and the supernatant-dependent inhibition of both K cell ADCC activity and lymphocyte-target cell conjugate formation. Together these data suggest that A-neutrophils suppress K cell function through a protease-mediated impairment of the FcR binding capacity. The results provide evidence that human neutrophils are endowed with mechanisms to regulate K cell ADCC activity.
In vitro restoration of polyclonal hematopoiesis in a chronic myelogenous leukemia after in vitro treatment with 4- hydroperoxycyclophosphamide
Authors: Degliantoni G , Mangoni L , Rizzoli V 
Published: October 2019

Abstract
Bone marrow cells of a 45-year-old female with Philadelphia chromosome (Ph1)-positive, early-phase chronic myelogenous leukemia (CML), who was heterozygous for the glucose-6-phosphate dehydrogenase (G6PD) locus, were pretreated in vitro with 4-hydroperoxycyclophosphamide (4-HC) and tested for G6PD activity in several colony formation assays and for karyotypic abnormalities. All cells within the mixed (CFU-GEMM), the erythroid burst (BFU-E), and the granulocyte-macrophage (CFU-GM) colonies expressed type A and type B G6PD activity and a normal karyotype, whereas untreated cells expressed type A G6PD and the Ph1 chromosome. This reversal of G6PD activity type and the disappearance of the Ph1 chromosome in colonies grown from 4-HC-treated cells indicate that this cytotoxic agent spares a residual normal stem cell population in bone marrow cells of early-phase CML patients. This finding, in turn, suggests a therapeutic approach in CML based on in vitro chemotherapy of autologous bone marrow grafts.
In vitro expansion and analysis of cloned cytotoxic T cells derived from patients with chronic T gamma lymphoproliferative disorders
Authors: van de Griend RJ , Bolhuis RL 
Published: October 2019

Abstract
Patients with T gamma lymphocytosis are a heterogeneous group, clearly distinguishable from other patients with chronic T lymphoproliferative disorders and usually without proven malignancy. We have attempted in vitro cloning of lymphocytes from three patients with an expansion of phenotypically and functionally different types of T gamma cells. One had T3+ B73.1+ T4 T8+ OKM1+ T gamma cells exerting antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) cell cytotoxicity; another had T3+B73.1-T4-T8+OKM1-ADCC+NK- and a third had T3-B73.1+T4-T8- OKM1+ADCC+NK+ cells. On morphological characterization, most of the mononuclear cells of these patients resembled large granular lymphocytes (LGLs). Although lymphocytes of these patients showed almost no proliferative response capacity after stimulation with mitogens, they shared the capacity to proliferate after stimulation with Epstein-Barr virus-transformed lymphoblastoid B (B-LCL) feeder cells. Stable clones were established by this procedure. Clones from patient 3 exerted cytolytic activity against a broad spectrum of tumor cell lines, including fresh biopsy specimens of melanoma tumor target cells. All of these clones (termed activated killer [AK] cells) had the surface phenotype T3-, T4-, T8- or +, HNK1-, OKM1-, Lyt3+, WT1+ and showed ADCC in addition to AK cell cytotoxicity. Most of them were B73.1+ and expressed IgG-Fc receptors. They most likely belong to the T cell lineage, since they express IL2 receptors as recognized by the Tac antibody and did not bind monoclonal antibodies directed against monocytes or granulocytes. Thus lymphocytes with the functional and phenotypical characteristics of T gamma cells can be cloned and expanded in vitro from the peripheral lymphocytes of these patients by using the appropriate stimulus. Our results indicate that, of the heterogeneous population of NK cells, the T3- cells are more rapidly expanded than T3+ subsets. It is discussed whether or not our culture system might selectively induce proliferation in “normal” T cells rather than aberrant ones.
Rapid changes in surface antigen expression by blood monocytes cultured in suspension or adherent to plastic
Authors: Triglia T , Burns GF , Werkmeister JA 
Published: October 2019

Abstract
Cell surface antigens used as markers of in vivo differentiation may not be stable on monocytes maintained under different conditions of in vitro culture. Monocytes were isolated from blood by centrifugation over Percoll or by adherence to plastic dishes, and the cells cultured in suspension or as adherent monolayers. Initially, monocytes obtained by both methods were similar in size, morphology, and surface antigen expression detected with the antimonocyte monoclonal antibodies OKM1, FMC17, PHM2, and PHM3. After culture, cells maintained in suspension were predominantly small, whereas those adherent to plastic rapidly increased in size; however, cytochemical staining for nonspecific esterase and acid phosphatase showed increased enzymic activity by monocytes in both systems, possibly reflecting increased cell maturation. The most striking difference was a substantial loss of FMC17 antigen by most monocytes within four hours in suspension culture, as compared with a qualitative and quantitative increase in expression by plastic adherent cells within two hours. These changes occurred even if the cells were first reacted with lipopolysaccharide. Monocytes taken from suspension culture and allowed to adhere to plastic rapidly synthesized the antigen, a process inhibited by cycloheximide, and conversely, cells removed from plastic progressively displayed decreased FMC17 antigen expression when transferred to suspension culture. No functional role in adherence or phagocytosis has been found for the FMC17 antigen. The results suggest that antigen expression may depend as much on the physical state of the cells as on apparent activation or maturation events.
Human spleen cell generation of factors stimulating human pluripotent stem cell, erythroid, and myeloid progenitor cell growth
Authors: Fabian I , Douer D , Levitt L , Kletter Y , Greenberg PL 
Published: October 2019

Abstract
Mitogen-stimulated murine spleen cells produce humoral substances capable of supporting murine hematopoiesis and pluripotent stem cell proliferation in vitro. Thus, we evaluated conditioned media generated by human spleen cells (SCM) in the presence or absence of mitogens for factors stimulatory for human pluripotent (CFU-GEMM), erythroid (BFU- E), and myeloid (CFU-GM) precursors. Two and one half percent to 10% SCM stimulated proliferation of all three types of precursor cells from nonadherent buoyant human marrow target cells. Mitogen-stimulated SCM augmented CFU-GM (175% to 225%), whereas CFU-GEMM and BFU-E growth was essentially unchanged. Cell separation procedures used to determine which cells provided these microenvironmental stimuli indicated that nonadherent mononuclear spleen cells provided the bulk of the CSF-GM, whereas adherent cells (95% nonspecific esterase + monocyte- macrophages) and nonadherent cells provided similar proportions of CSF- mix and erythroid burst-promoting activity (BPA). The nonadherent cells generating high levels of CSF-mix, BPA, and CSF-GM were predominantly Leu-1-negative, ie, non-T, cells. In the presence or absence of mitogens, SCM was a more potent source (1.3- to 3.8-fold) than peripheral leukocyte CM of the growth factors for the three progenitor cell types. Specific in situ cytochemical stains for analyzing morphology of myeloid colonies demonstrated that SCM stimulated the proliferation of the same types and proportions of colonies as human placental CM, suggesting that these CMs may contain similar CSF-GMs. These data show the contribution of spleen cell subsets to the generation of hematopoietic growth factors and the responsiveness of these cells to various mitogenic stimuli.