bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment of refractory adult lymphoblastic leukemia (ALL) with 4'(9- acridinylamino) methanesulfon-M-anisidide (AMSA)
Authors: Arlin ZA , Fanucchi MP , Gee TS , Kempin SJ , Mertelsmann R , Young CW , Clarkson BD
Published: October 2019

Twenty-four adults with ALL were treated with AMSA alone or in combination. Twenty-two were treated at time of relapse and two patients after failing primary induction therapy. All had been treated with anthracyclines prior to receiving AMSA. Of the 22 patients with ALL in relapse, 4 achieved a complete remission. Two of these patients have relapsed while receiving maintenance chemotherapy; one died 1 mo after achieving remission due to the occurrence of cholycystitis in the setting of pancytopenia and one patient underwent bone marrow transplantation and is in remission at 8 mo after the second remission. Both patients who failed primary induction therapy remain in remission at 11 and 36 mo, respectively. The use of AMSA should be considered for patients with ALL who fail primary induction as well as those whose leukemia becomes resistant to conventional agents.
Thrombin-induced secretion of serotonin from platelets can occur in seconds
Authors: Gear AR , Burke D
Published: October 2019

The platelet release reaction was studied by a new quenched-flow approach. Platelets labeled with 14C-serotonin were reacted for short times (up to 5 sec) with thrombin and then quenched with glutaraldehyde or paraformaldehyde. Serotonin secretion began within 1 sec and was nearly complete by 4 sec. Aggregation recorded by a resistive-particle counter was similarly fast. Therefore, the quenched-flow system reveals that serotonin secretion can occur more rapidly than estimated in earlier studies.
Prognostic significance of terminal transferase activity in childhood acute lymphoblastic leukemia: a prospective analysis of 164 patients
Authors: Hutton JJ , Coleman MS , Moffitt S , Greenwood MF , Holland P , Lampkin B , Kisker T , Krill C , Kastelic JE , Valdez L , Bollum FJ
Published: October 2019

Whether the level of terminal deoxynucleotidyl transferase (TdT) activity in mononuclear cells from bone marrow and peripheral blood has prognostic significance has been analyzed prospectively in 164 children with T and non-T, non-B marked acute lymphoblastic leukemia (ALL). TdT was measured at diagnosis to assess its value as a predictor of duration of remission and length of survival. It was measured repeatedly during remission to assess whether it could predict relapse. Ninety-seven percent of the children achieved a complete remission of their disease, and 40% relapsed during the study. The level of TdT activity in blasts at diagnosis varied 1000-fold from patient to patient. There was no statistically significant relationship between TdT activity in cells at diagnosis and the achievement of complete remission, the duration of remission, or length of survival. TdT activity was significantly increased in the bone marrow of 65% of patients at the time of marrow morphological relapse, but was rarely increased in marrow from patients with isolated testicular or central nervous system relapse. Wide fluctuations in TdT activity were characteristically seen in mononuclear cells from the marrow and peripheral blood of patients with ALL at all stages of their disease. An isolated high value of TdT activity in the bone marrow or peripheral blood cannot be taken as evidence of impending relapse. Quantitative measurements of TdT activity alone on mononuclear cells from bone marrow and peripheral blood are helpful in differential diagnosis, but cannot guide therapy of children with ALL.
A defect in the oxidative metabolism of human polymorphonuclear leukocytes that remain in circulation early in hemodialysis
Authors: Cohen MS , Elliott DM , Chaplinski T , Pike MM , Niedel JE
Published: October 2019

Human granulocytes harvested from uremic volunteers 15 min after the initiation of dialysis (at the nadir of neutropenia) were compared to predialysis controls. These intradialysis cells had a significant defect in peak luminol-enhanced chemiluminescence in response to opsonized zymosan, f-Met-Leu-Phe, and phorbol myristate acetate relative to predialysis control cells from the same patients. This defect could not be explained by a decrease in PMN myeloperoxidase concentration. H2O2 secretion by intradialysis cells (2 patients) was also depressed relative to predialysis controls. The ability to perform an independent function, orientation (polarization), was normal in both pre- and intradialysis cells relative to control. Whereas 125I-labeled formyl peptide binding studies demonstrated identical values for affinity and receptor number for predialysis and normal control cells, intradialysis cells displayed a 27% decrease in receptor number. This decrease in available receptor number. This decrease in available receptors may be related to the decreased chemiluminescence observed in response to f-Met-Leu-Phe. Furthermore, the results are consistent with the hypothesis that a defective PMN population remains in the circulation during the neutropenia of hemodialysis.
Normal human pluripotential and committed hematopoietic progenitors do not express the p24 antigen detected by monoclonal antibody BA-2: implications for immunotherapy of lymphocytic leukemia
Authors: Ash RC , Jansen J , Kersey JH , LeBien TW , Zanjani ED
Published: October 2019

Analysis of surface antigenic determinants of hematopoietic progenitor cells has relevance both to basic biologic study of cell differentiation and to potential clinical application in the diagnosis and treatment of hematologic neoplasia. The production and characterization of monoclonal antibody BA-2 by immunization with a pre- B-ALL cell line has been reported previously. In this study we utilized complement-dependent cytotoxicity and rosette-separation with antibody indirectly coupled to ox RBC to determine if the antigen (p24) recognized by the antibody BA-2 is represented on human pluripotential (CFU-GEMM) or committed hematopoietic progenitors (CFU-GM, BFU-E, CFU- E). BA-2 showed no reactivity with normal hematopoietic progenitors by either method. In contrast, BA-2 exhibited potent complement-mediated cytotoxicity for selected ALL-derived cell lines. These results show that normal human hematopoietic progenitors do no express antigenic sites represented on ALL cells that are recognized by BA-2 and suggest that this monoclonal antibody may serve as a potent and specific agent for treatment of lymphocytic leukemia, perhaps most useful in ex vivo marrow conditioning for autologous bone marrow transplantation.
The role of the spleen in regulating the plasma levels of factor VIII-- von Willebrand's factor after DDAVP
Authors: Garcia VV , Coppola R , Mannucci PM
Published: October 2019

Organ transplantation and perfusion studied indicate that the spleen plays an important role in the regulation of plasma levels of factor VIII-von Willebrand's factor (FVIII-vWF). To better understand the mechanisms that regulate the FVIII-vWF increases after infusion of 1- deamino-8-D-arginine vasopressin (DDAVP), we have measured factor VIII coagulant activity (FVIII:C) and antigen (FVIII:CAg) and von Willebrand's factor antigen (vWF:Ag) and ristocetin cofactor (vWF:RCof) in 9 asplenic subjects with normal baseline concentrations, in 7 asplenic subjects with high concentrations, and in 14 normal controls with intact spleens. In “normal” aasplenics, all the FVIII-vWF-related measurements increased significantly over baseline values, indicating that responsiveness to DDAVP is not abolished by splenectomy. The maximal vWF:Ag and vWF:RCof responses were no different from those of normal controls, suggesting that DDAVP releases vWF from storage sites other than the spleen. The FVIII:C response was significantly lower than in normal controls, but FVIII:CAg did not differ, making FVIII:CAg higher than FVIII:CAg in “normal” asplenics. These findings suggest that the spleen, rather than being a storage site for FVIII, is the organ in which a partially inactive form of FVIII acquires full coagulant activity. In “high” asplenics, all the FVIII-vWF-related measurements increased less than in “normal” splenics, indicating that long-term elevations of plasma concentrations of FVIII-vWF are accompanied by decreased release from those storage pool(s) mobilized by DDAVP.
Generation of CFU-C suppressor T cells in vitro. III. Failure of mitogen-primed T cells from patients with chronic granulocytic leukemia to inhibit the growth of normal CFU-C
Authors: Frassoni F , Bacigalupo A , Podesta M , Van Lint MT , Piaggio G , Marmont A
Published: October 2019

T lymphocytes were derived by E rosetting from the peripheral blood (PB) and bone marrow (BM) of 15 patients with chronic granulocytic leukemia (CGL) in the chronic phase of their disease. T cells were also obtained from 12 healthy individuals. T cells were incubated overnight either in culture medium (RPMI) or RPMI plus pokeweed mitogen (PWM). The supernatants were then recovered and the cells washed in fresh RPMI. T cells from normal donors and from CGL patients were then cocultured with normal allogeneic marrow cells grown in soft agar for CFU-C colony formation. Target marrow cells were also grown in agar in the presence of T-derived supernatants. The results of this study can be summarized as follows. (1) Normal PB and BM T cells efficiently suppressed autologous and allogeneic CFU-C growth after PWM stimulation. (2) T cells derived from peripheral blood or marrow of CGL patients failed to inhibit CFU-C growth, whether pretreated with PWM or not. (3) The supernatants of PWM-treated normal T cells strongly inhibited CFU-C colony formation, whereas the supernatants of PWM- treated CGL T cells had no CFU-C/suppressor activity. These data indicate that T cells from CGL patients cannot be primed to become CFU- C suppressor cells after PWM: stimulation in vitro and cannot release a soluble inhibitor of granulopoiesis produced by PWM-primed normal T cells.
Human platelet cytoskeletons: specific content of glycolipids and phospholipids
Authors: Schick PK , Tuszynski GP , Vander Voort PW
Published: October 2019

The lipid composition of platelet cytoskeletons was analyzed. Triton X- 100 (0.5%) was used to prepare cytoskeletons from thrombin-treated platelets. The lipid/protein ratio of platelet cytoskeletons was 0.260 and the phospholipid/protein ratio was 0.177, which were comparable to the ratios present in platelets. However, there was a selective enrichment of platelet lipids in platelet cytoskeletons. Only 2 of the 5 major platelet phospholipids were detected. About 14% platelet sphingomyelin and 2% platelet phosphatidylcholine were present in platelet cytoskeletons. Only 1 of the 4 platelet neutral glycolipids, trihexosyl ceramide, was detected and was about 7% of that in intact platelets. Two percent of platelet hematoside, the predominant ganglioside in platelets, was found in cytoskeletons. Six percent of platelet cholesterol was present in platelet cytoskeletons, while no other neutral lipid could be detected. The study demonstrates that the lipid/protein ratio of platelet cytoskeletons is similar to that in platelets, but the composition of cytoskeleton lipids is specific and distinctly different from that in platelets. The selective glycolipid and phospholipid composition of cytoskeletons may be important for cytoskeleton and platelet function.
Heterogeneous ontogeny of erythropoiesis after bone marrow ablation and allogeneic bone marrow grafting
Authors: Winkler KJ , Rea CD , Rahbar S , Hill LR , Blume KG
Published: October 2019

Thirty-two patients who underwent bone marrow transplantation for hematologic malignancies were studied for hemoglobin-F as an indicator for fetal erythropoiesis. Two different patterns of response were noted. One group of patients who had an elevated HbF level prior to marrow grafting later showed a marked reactivation of HbF synthesis, whereas the other group of patients who had normal HbF levels prior to transplantation failed to exhibit such an increase. This phenomenon occurred independently of ages of marrow graft donors or recipients, the type of underlying hematologic malignancy, or remission induction therapy prior to preparation for transplantation, pretransplant hemoglobin levels, transfusion with red blood cells, red cell volume, and production of reticulocytes.
Immunoreactive calcitonin production by a human promyelocytic leukemia cell line HL60
Authors: Oscier DG , Hillyard CJ , Arnett TR , MacIntyre I , Goldman JM
Published: October 2019

Using a sensitive radioimmunoassay,we detected human immunoreactive calcitonin in cell extracts and in cell-exposed media of the HL60 cell line derived originally from a patient with acute promyelocytic leukemia. The cell extract was chromatographed on a reverse-phase high- pressure liquid chromatography system. Radioimmunoassay of the fractions showed that the immunoreactive calcitonin was heterogeneous but had peaks corresponding to those of synthetic human calcitonin monomer and its sulphoxide. We have previously shown that levels of immunoreactive calcitonin are elevated in the plasma of the majority of patients with acute and chronic myeloid leukemias. Our studies with the HL60 cell line add further support to the concept that leukemic cells can synthesize immunoreactive calcitonin “ectopically.”
A radiolabeled antiglobulin test for crossmatching platelet transfusions
Authors: Kickler TS , Braine HG , Ness PM , Koester A , Bias W
Published: October 2019

Despite the use of HLA-matched platelets for alloimmunized recipients, transfusion failures occur. In order to reduce these failures, we investigated the use of a radiolabeled antiglobulin technique for platelet crossmatching. The principle of the test is that of an indirect Coombs test using 125I labeled goat anti-human IgG. Incompatibility is determined by calculating a radioactivity antiglobulin test (RAGT) index. Using this technique, we performed 89 crossmatches on 19 leukemic or aplastic patients who were refractory to random donor platelets and receiving varying degrees of HLA-matched platelets. Effectiveness of the transfusion was assessed from the posttransfusion corrected platelet count increment (CCI) determined at 1 and 20 hr. When the RAGT index was 1.9 or less, the mean CCI at 1 lhr was 17,570 +/- 7003/cu mm, n = 55. When the RAGT index was 2.0 or greater, the mean CCI was 4237 +/- 4100/cu mm, n = 34. At 20 hr when the RAGT index was 1.9 or less, the mean CCI was 8722 +/- 3143/cu mm, n = 33, and when the index was 2.0 or greater, the mean CCI was 571 +/- 1286/cu mm, n = 23. Using this technique, one false negative resulted. Nine positive crossmatches with good increments at 1 hr were found; at 20 hr, however, the survival of these units was zero. These data suggest that this method is a useful adjunct in the selection of platelets in the refractory patient.
An investigation of 2'-deoxycoformycin in the treatment of cutaneous T- cell lymphoma
Authors: Grever MR , Bisaccia E , Scarborough DA , Metz EN , Neidhart JA
Published: October 2019

We have investigated the use of 2′-deoxycoformycin (DCF), a potent inhibitor of adenosine deaminase (ADA), in the treatment of 4 patients with advanced mycosis fungoides (MF). Since DCF has demonstrated an adverse effect in vitro and in vivo on the survival of leukemia T-cell lines, it appeared reasonable to examine its effect in patients with advanced cutaneous T-cell lymphoma. A total of 8 courses of DCF were given to the 4 patients. Since this study was part of an ongoing phase I investigation, each patient received a fixed dose (varying from 4 mg/sq m to 10 mg/sq m daily for 3 consecutive days) on a 28-day schedule. One patient had reversible renal insufficiency. Three patients had reversible myelosuppression. Two patients had a complete remission of disease for 7+ and 9+ mo. respectively. Two additional patients had partial remissions for 4 and 9 mo, respectively. We concluded that effective antitumor activity in advanced MF can be achieved with DCF at doses that may not be associated with prohibitive toxicity. We would encourage further investigation of this agent in patients with advanced cutaneous T-cell lymphoma.
Differential excretion of modified nucleosides in adult acute leukemia
Authors: Heldman DA , Grever MR , Trewyn RW
Published: October 2019

Excretion of modified nucleosides in urine was measured in 23 adults with acute leukemia to determine correlation of nucleoside excretion with disease activity. In addition, differences in excretion between patients with acute lymphoblastic leukemia (ALL) and patients with acute myeloid leukemia (AML) were established. Six modified nucleosides were resolved and quantitated by reversed-phase high-performance liquid chromatography (HPLC). Patients with ALL at initial diagnosis or in relapse had significantly higher concentrations of 1-methylinosine and N2,N2-dimethylguanosine in their urine compared to patients in remission (p less than 0.01, p less than 0.05, respectively). One patient with ALL was followed with serial nucleoside determinations over a period of 18 mo; nucleoside excretion correlated closely with disease activity. Nucleoside excretion in patients with AML did not change significantly with disease activity. Considering only those patients at initial diagnosis or in relapse, excretion of 1- methylinosine and N2,N2-dimethylguanosine was significantly higher in ALL than in AML (p less than 0.01, p less than 0.05, respectively). Thus, urinary excretion of 1-methylinosine and N2,N2-dimethylguanosine by adults with acute leukemia may prove to be valuable clinically in following disease activity in patients with ALL and in distinguishing patients with ALL from those with AML.
Isoelectric focusing of human von Willebrand factor in urea-agarose gels
Authors: Fulcher CA , Ruggeri ZM , Zimmerman TS
Published: October 2019

An analytical technique has been developed for the isoelectric focusing (IEF) of plasma von Willebrand factor (vWF) in agarose gels containing urea. Under these conditions, vWF freely enters the gel and focuses without artifact. The focused vWF is visualized by staining fixed gels with 125I-labeled affinity-purified heterologous antibody. Utilizing a pH gradient of 5.0–6.5, normal vWF in plasma or purified preparations focuses into at least three bands with apparent isoelectric points (pI) between pH 5.7 and 5.9. A reproducible difference in the IEF pattern of vWF has been established between normal plasmas and those of individuals with variant von Willebrand's disease (vWd) type IIA and type IIB. In type IIA, vWF has a distinctly lower pI than normal. This difference may be related to the presence of smaller vWF multimers in IIA plasma because forms of vWF of corresponding size contained in normal cryoprecipitate supernatant have a similar pI. Type IIB von Willebrand factor has a pI intermediate between normal and IIA. Neuraminidase treatment of plasma samples before IEF results in an increase in pI in normal, type IIA, and type IIB vWF. The data suggest that none of the 16 type IIA and 9 IIB plasmas studied here contain significantly decreased amounts of sialic acid.
In vitro studies of megakaryocytopoiesis in thrombocytotic disorders of man
Authors: Gewirtz AM , Bruno E , Elwell J , Hoffman R
Published: October 2019

Increased numbers of bone marrow megakaryocytes and thrombocytosis are frequently observed in patients with myeloproliferative disorders (MPD). Increased marrow megakaryocytes and thrombocytosis are also noted in a variety of inflammatory and neoplastic disease leading to the phenomenon of reactive thrombocytosis (RT). The pathogenesis of this finding remains incompletely understood. Using methodology developed in our laboratory, we investigated the causative role of megakaryocyte colony-stimulating activity (Meg-CSA) in generating this phenomenon. We also examined the cloning efficiency of colony-forming units-megakaryocyte (CFU-M) and their responsiveness to an exogenous source of Meg-CSA in patients with these diseases. The results of our investigations suggest that: (1) increased production of Meg-CSA is not responsible for the megakaryocyte hyperplasia and thrombocytosis noted in these patients; (2) the intrinsic stem cell defect described in MPD appears to affect the CFU-M of these patients as well, resulting in an effective expansion of the CFU-M pool with consequent megakaryocyte hyperplasia and thrombocytosis; (3) the CFU-M of patients with MPD remain responsive to an exogenous source of Meg-CSA, suggesting that this megakaryocyte hyperplasia may not be entirely autonomous of its effects; and (4) the CFU-M pool in RT is normal both in size and responsiveness to Meg-CSA, suggesting that in these disorders, the stimulus leading to megakaryocyte hyperplasia and thrombocytosis is active at the post-CFU-M level of megakaryocyte differentiation.
Evidence of the existence of a factor that induces C3 receptors on bone marrow cells
Authors: Calcagno M , Rios B , Fragoso A , Arciga MA , Cabrera G , Torres R , Weiss-Steider B
Published: October 2019

The existence of a molecule responsible for the induction of C3R receptors (C3R) on murine bone marrow cells (C3R inducer, C3RI) is demonstrated in lung conditioned media. The inducer of Fc receptors on murine bone marrow cells (FcRI) and the colony-forming factor MGI (macrophage and granulocyte inducer) are also identified from the same source. The three inducing factors are shown by molecular sieving chromatography and by isoelectric focusing in granulated gel to be three different molecules. The molecular weights estimated were of 35,000, 22,000, and 10,500 for C3RI, MGI, and FcRI, respectively, with isoelectric pH values of 3.9, 6.6, and 7.6 for the same molecules. The C3RI is assumed to be of protein nature, because the molecule lost its biologic activity after trypsin or heat treatment. In the two techniques used, the three peaks of activity were completely separated, suggesting that the induction of C3 or Fc receptors and colony formation are mediated by different molecules and thus regulated independently at the cellular level.
Dose-dependent antithrombotic effect of warfarin in rabbits
Authors: Gitel SN , Wessler S
Published: October 2019

One-hundred and fifty-one rabbits, divided into controls and animals treated with varying daily doses of warfarin, were subjected to the stasis assay, and the amount of thrombosis quantitated after intravascular coagulation was initiated either by activated factor X or tissue thromboplastin. Following 8–10 days of warfarin administration, there was a significant dose-dependent decrease in the vitamin-K- dependent coagulation factors paralleled by an increase in the prothrombin time ratio. Whether thrombosis was initiated by activated factor X or tissue thromboplastin, there was, with increasing drug dose, a progressive increase in the inhibition of stasis thrombosis. This significant antithrombotic effect occurred even when the vitamin-K- dependent coagulation activities were at a mean value of 50%.
Studies of T-lymphocytes in preleukemic disorders and acute nonlymphocytic leukemia: in vitro radiosensitivity, mitogenic responsiveness, colony formation, and enumeration of lymphocytic subpopulations
Authors: Knox SJ , Greenberg BR , Anderson RW , Rosenblatt LS
Published: October 2019

Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and lymphocyte colony formation (CFU-L) from whole blood were measured following in vitro x-irradiation. Lymphocytes from patients with myelodysplastic disorders, acute nonlymphocytic leukemia, and patients at increased risk for leukemia because of their primary disease and/or cytotoxic therapy were found to be significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals. Cloning efficiencies and mitogenic responsiveness of patient lymphocytes were significantly depressed as compared to normal values. Using monoclonal antibodies to specific surface markers, quantitative abnormalities in lymphocytic subpopulations from myelodysplastic patients also were observed. These findings are suggestive of a defect at the T-cell level that may directly or indirectly affect hematopoiesis.
Monoclonal antibody studies in non-Hodgkin's lymphoma
Authors: Aisenberg AC , Wilkes BM , Harris NL
Published: October 2019

The cell lineage of suspensions prepared from 85 non-Hodgkin's lymphomas was investigated with a panel of 10 monoclonal antibodies and conventional surface marker techniques. Surface immunoglobulin, assessed with specific heteroantisera, proved to be the most useful characteristic and defined the clonal character and B-cell lineage of 63 specimens: almost all nodular lymphocytic (21 of 22) and diffuse lymphocytic (11 of 13) lymphomas, most diffuse histiocytic (29 of 33) and diffuse mixed (2 of 2) lymphomas, and a few nodular mixed (2 of 12) and nodular histiocytic (0 of 3) lymphomas. Monoclonal antibodies provided useful ancillary surface marker criteria. Thus, positivity with OKT1 (which detects both thymic and peripheral T cells) in the absence of reactivity with monoclonal antisera, which detect only peripheral T cells (OKT3, OKT4, OKT8, and OKT11), was seen only in diffuse lymphocytic lymphoma of B lineage. Ia-like antigen could be demonstrated in all B-cell lymphocytic lymphomas and most B-cell diffuse histiocytic lymphomas. Approximately one-half of diffuse histiocytic lymphomas also reacted with OKT9, which detects the transferrin receptor, while few lymph nodes involved by other conditions displayed this reactivity. Most diffuse histiocytic lymphomas and many non-Hodgkin's lymphomas of other subtypes reacted with OKT10, an antiserum that detects an antigen on replicating lymphoid cells. The lineage of approximately one-fourth of the lymphoma suspensions was not resolved conclusively: In most of these, T lymphocytes predominated with a normal proportion of inducer-helper (OKT4) and cytotoxic-suppressor (OKT8) cells. The inability to establish the clonal character of T-cell proliferation in cell suspensions remains an obstacle to completely defining the lineage of non-Hodgkin's lymphomas.
Myeloproliferative syndrome induced by MPSV in DBA/2 mice: presence of a mixed-colonies promoting activity (MPA) in the spleen
Authors: Le Bousse-Kerdiles MC , Smadja-Joffe F , Klein B , Jasmin C , Comisso M , Ostertag W
Published: October 2019

The myeloproliferative syndrome induced by the myeloproliferative sarcoma virus (MPSV) in DBA/2 mice stimulates the proliferation of pluripotent hemopoietic stem cells (HSC) and of progenitors committed toward granulomacrophagic and erythroid cell lines. This stimulation may result from a direct effect of the MPSV on HSC or from an indirect effect via locally secreted factors. Normal isogenic bone marrow cells were incubated in the mixed colony-forming unit system in semisolid medium supplemented with conditioned media obtained after incubating neoplastic spleen cells for 3 days at 37 degrees C. These spleen conditioned media contain an activity that is physically separable from MPSV by ultracentrifugation and which, in the presence of a very low quantity of erythropoietin, can induce in vitro the proliferation and differentiation of pluripotent HSC, detected by this Mix-CFU technique. We termed this activity mixed-colonies promoting activity (MPA). These results suggest that the hyperplasia of the nonlymphoid hematopoietic system in the neoplastic spleen results from an indirect effect of the MPSV on pluripotent HSC via locally secreted factors.
Characterization of normal peripheral blood lymphocyte colony-forming cells: cell cycle status, surface markers, and cellular growth requirements
Authors: Taetle R , To D , Caviles A , Norby SW , Mendelsohn J
Published: October 2019

We performed a series of studies to further clarify the nature of lymphocyte colony-forming cells (CFC) from normal peripheral blood. Mononuclear cells were separated into E-rosette-enriched (E+) and E- rosette-depleted (E-) populations and cultured in methylcellulose with conditioned media and irradiated mononuclear cells. Linear plating relationships were obtained with plating efficiencies of 0.26% +/- .02% (mean +/- SE) for E+ CFC and 0.18% +/- .02% for E- CFC. Cells in E+ colonies were T lymphocytes and in E- colonies were B lymphocytes as determined by cell surface marker analysis. Using the thymidine suicide technique, approximately one-half of CFC were found to be in cycle at any moment, and plating efficiencies and cell cycle status of E+ CFC were not changed by preincubation with PHA in liquid culture for 48 hr. Using antibody complement-mediated cytotoxicity, E+ CFC were found to be T101+, OKT3+, and Ia-, while E- CFC were OKT3- and Ia+. Using monocyte-depleted populations obtained by sedimentation at unit gravity, lymphocyte colony growth was absent in monocyte-depleted fractions, and optimal growth occurred with 40% monocytes in culture. In contrast to some previous studies, we find that lymphocyte CFC originate from a small, cycling population of cells bearing mature T or B lymphocyte markers. Entry into cell division, however, does not confer colony-forming capacity on lymphocytes. Monocytes are critical to growth of E+ CFC, and cultures severely depleted of monocytes would not be expected to form colonies.
Characterization of human megakaryocyte colony-stimulating factor in the urinary extracts from patients with aplastic anemia and idiopathic thrombocytopenic purpura
Authors: Kawakita M , Ogawa M , Goldwasser E , Miyake T
Published: October 2019

Using procedures that were effective in the purification of human urinary erythropoietin (Epo), we attempted initial purification of megakaryocyte colony-stimulating factors (CSF) in urinary extracts from patients with aplastic anemia (AA) and idiopathic thrombocytopenic purpura (ITP). Comparison of colony stimulation by purified human Epo and crude urinary extracts revealed: (1) that the pure Epo augments megakaryocyte colony formation in culture and (2) MEG-CSF activity is also present in materials other than Epo in the crude urinary extracts from the two types of patients. Similar to purification of Epo, ethanol precipitation and sulfopropyl-Sephadex chromatography provided twofold and threefold increases in the specific activity of MEG-CSF, respectively. In contrast to Epo, however, significant inactivation of MEG-CSF activity was seen with phenol treatment. The elution profile of MEG-CSF seen on hydroxylapatite chromatography of urinary extracts was different from that of Epo. These data provided a basis for initial steps for purification of MEG-CSF and support the notion that MEG-CSF is distinct from Epo.
Effects of 12-0-tetradecanoylphorbol-13-acetate (TPA) on the proliferation of granulocyte-macrophage colony-forming cells
Authors: Burgess AW , Nicola NA
Published: October 2019

It has been suggested that 12–0-tetradecanoylphorbol-13-acetate (TPA) may stimulate the proliferation of granulocyte-macrophage (GM) colony- forming cells (CFC) via the GM colony-stimulating factor (CSF) receptor. GM-CFC in unfractionated mouse bone marrow and light density fetal liver (LDFL) cells were induced by TPA to form colonies in the absence of exogenously added GM-CSF. The colonies induced by TPA (10(- 8)M) were smaller than normally seen with maximal concentrations of GM- CSF, and less than 30% of the GM-CFC formed colonies in the presence of TPA. The number of colonies stimulated by TPA in the absence of GM-CSF was dependent on the number of cells plated. When fewer than 10,000 bone marrow cells or 3000 LDFL cells were plated in the 1-ml semisolid agar cultures, no colonies were stimulated by the TPA. Similarly, GM- CFC purified from the LDFL cells stimulated with TPA did not form colonies. However, when the fetal liver accessory cells (macrophages) were recombined with cell-sorter-purified GM-CFC, colony formation was again observed in the presence of TPA (10(-7)-10(-8) M). The number of colonies formed from the CFC was dependent on the number of accessory cells present, suggesting that the macrophages were induced by TPA to produce CSF. Although the purified GM-CFC required CSF for proliferation, TPA (10(-8) M) increased (5–10-fold) the sensitivity of the GM-CFC to GM-CSF. These observations indicate that TPA does not stimulate GM-CFC proliferation directly, but rather by inducing GM-CSF production by accessory cells and by increasing the responsiveness of GM-CFC to GM-CSF.
Granulocytosis associated with tumor cell production of colony- stimulating activity
Authors: Hocking W , Goodman J , Golde D
Published: October 2019

A patient with metastatic soft tissue sarcoma presented with a leukemoid reaction. The elevated white blood cell count was due to an increase in bands and mature segmented neutrophils. The degree of granulocytosis correlated with the tumor burden. There was no evidence of superimposed infection and the degree of bone marrow involvement by metastatic tumor was minimal. A cell line derived from the sarcoma produces granulocyte-macrophage colony-stimulating activity (CSA) in vitro. Although CSA could not be detected in serum, the findings in this patient suggest that the leukemoid reaction was due to tumor production of CSA.
Regulation of hematopoiesis II: the role of polyamine inhibition on helper or suppressor influences of the thymus
Authors: Sharkis SJ , Luk GD , Collector MI , McCann PP , Baylin SB , Sensenbrenner LL
Published: October 2019

We have previously suggested in murine model systems, that two cell subpopulations with differing proliferative capacity, from the thymus, modify the growth of erythroid progenitor cells in vitro. In order to further characterize these populations, we have specifically inhibited polyamine biosynthesis; this pathway is essential for the process of cell replication. Thus, alpha-difluoromethyl ornithine (DFMO) was used to block the conversion of ornithine to putrescine, the first and rate- limiting step in polyamine biosynthesis. We observed a threefold increase in hematopoietic progenitors (CFU-S and CFU-E) from bone marrow in animals treated with DFMO. We further examined the effect of DFMO on accessory “helper” and “suppressor” cells from the thymus and observed an increase in helper activity with an elimination of suppressor activity. All of these effects of DFMO were specific for inhibition of polyamine biosynthesis, since simultaneous addition of the depleted biosynthetic product, putrescine, restored suppressor activity. We conclude that polyamine biosynthesis is required acutely for accessory cell regulation of hematopoiesis.
Cholesterol and mevalonic acid are independent requirements for the in vitro proliferation of human bone marrow granulocyte progenitor cells: studies using ML-236B
Authors: Hoffman PC , Richman CM , Larson RA , Yachnin S
Published: October 2019

ML-236B is a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, the key regulatory enzyme in the sequence that catalyzes the conversion of acetate to mevalonic acid in cholesterol biosynthesis. This compound caused marked inhibition of human bone marrow granulocyte progenitor cell (CFU-C) proliferation, the 50% inhibitory concentration (IHD50) being 2.0 X 10(6)M. Inhibition of colony formation was reversed by mevalonic acid but not by cholesterol. ML-236B also inhibited DNA synthesis and acetate incorporation into cholesterol in marrow mononuclear cells (IHD50 = 5.6 x 10(6)M and 3.2 x 10(7)M, respectively). No inhibition of mevalonate incorporation into cholesterol was observed. These results differ from those observed with 25-hydroxycholesterol, another inhibitor of HMG CoA reductase. The latter compound also inhibited CFU-C proliferation and cholesterol biosynthesis from acetate; inhibition of colony formation was reversed by cholesterol but not by mevalonic acid. In addition, 25- hydroxycholesterol inhibited cholesterol synthesis from mevalonic acid precursor. We conclude that: (1) ML-236B is a potent inhibitor of CFU-C proliferation, DNA synthesis, and cholesterol biosynthesis from acetate precursor in marrow mononuclear cells; (2) the effects of ML-236B are completely reversed by mevalonic acid but not by cholesterol, suggesting that mevalonic acid per se or one or more of its nonsterol products are critical for cell growth; (3) the inhibitory effects of 25- hydroxycholesterol on CFU-C proliferation and cholesterol biosynthesis are not solely a result of its inhibition of HMG CoA reductase, but are due in part to inhibition of enzymatic steps distal to mevalonic acid in the sterol synthetic pathway; and (4) mevalonic acid and cholesterol are independent requirements for CFU-C proliferation and differentiation in vitro.
Procoagulant activity of lymphocyte-macrophage populations in rabbits: selective increases in marrow, blood, and spleen cells during Shwartzman reactions
Authors: Rothberger H , Dove FB , Lee TK , McGee MP , Kardon B
Published: October 2019

The present experiments examine leukocyte procoagulant activity using mononuclear cell populations purified or enriched from rabbit bone marrow, blood, spleen, lymph node, thymus, and pulmonary alveoli. Cells from these six sites, obtained from control and endotoxemic animals and assayed without an intermediate culture step, were found to have procoagulant activity identified as tissue factor. Under control conditions, tissue factor activity was found to be at low levels in marrow and blood populations compared to median activities 3- and 11- fold higher in populations from spleen and lymph node, and 33- and 45- fold higher in thymus and alveolar populations. By contrast to respective controls, significantly increased amounts of tissue factor (35-, 15-, and 12-fold at median levels) were found in marrow, blood, and spleen populations from endotoxemic animals. The types of leukocytes in these latter three populations were morphologically and histochemically indistinguishable from respective controls, indicating that endotoxin induced increases of activity in cells with relatively low amounts under control conditions. Activity did not change significantly in lymph node, thymus, or alveolar populations after endotoxemia. These studies show that tissue factor is present in a range of leukocyte populations not previously reported to have procoagulant activity. In addition, the finding of widespread gains of tissue factor in the marrow-blood-spleen pool due to endotoxemia provides new evidence supporting the importance of leukocyte procoagulants in Shwartzman-like reactions.
The role of polyamine biosynthesis in hematopoietic precursor cell proliferation in mice
Authors: Niskanen E , Kallio A , McCann PP , Baker DG
Published: October 2019

Under the influence of a selective irreversible inhibitor of ornithine decarboxylase (ODC), DL-alpha-difluoromethylornithine (DFMO), early hematopoiesis was enhanced. In the bone marrow, the absolute number of cells that give rise to spleen colonies in lethally irradiated mice (CFU-S), granulocytic colonies in diffusion chambers in mice (CFU-DG), and granulocyte-monocyte colonies in agar in vitro (CFU-C) was increased 2–4 fold. This could be abrogated by administration of putrescine, confirming the association of the stimulatory effect with polyamine biosynthesis most likely via depression of ornithine decarboxylase activity and subsequent synthesis of putrescine. Analysis of cell cycle characteristics by 3H-TdR suicide technique demonstrated that the proportion of CFU-S, CFU-DG, and CFU-C in S-phase was significantly increased. Additionally, the stimulatory effect was reflected by enhanced colony formation in diffusion chambers implanted intraperitoneally in mice receiving DFMO. This could also be eliminated by treatment of the host animal with putrescine, again suggesting that polyamine biosynthesis plays an important role at the early stages of hematopoiesis in vivo. Effect of DFMO on colony formation in vitro (CFU- C) was inhibitory and not reversible with putrescine. It could be partially eliminated by aminoguanidine, which neutralizes diamine oxidase present in fetal calf serum used in the CFU-C assay. These data suggest that the effect of DFMO in vitro was nonspecific.
Production and regulation of interleukin-2 in human lymphoblastic leukemias studied with T-cell monoclonal antibodies
Authors: Venuta S , Mertelsmann R , Welte K , Feldman SP , Wang CY , Moore MA
Published: October 2019

Human leukemias are illnesses of hemopoietic stem cells that go through processes of self-replication and partial differentiation under the control of as yet largely unknown growth and differentiation factors. IL-2 is a powerful factor controlling proliferation of normal T cells. We report that acute lymphoblastic leukemias of T and non-B, non-T phenotypes produce a growth factor after mitogen stimulation. This factor is able to support the proliferation of human and murine IL-2- dependent cytotoxic cells, has a mol wt of 26,000 daltons by gel filtration, an isoelectric point of 6.6, and its biologic activity is inhibited by an anti IL-2 monoclonal antibody. This factor is, therefore, by all parameters studied very similar to IL-2 produced by normal lymphocytes. A recently developed monoclonal antibody, Pan T2, binds to normal T cells, renders T cells responsive to IL-2, and induces the release of IL-2, which in turn provides the second signal for T-cell proliferation. Mononuclear cells from acute lymphoblastic leukemia do not respond to the addition of this monoclonal antibody unless cocultured with irradiated Daudi cells. Since normal T cells do not require Daudi to produce IL-2 and since Daudi cells do not produce IL-2 under any conditions, we conclude that the cell responsible for IL- 2 production in acute lymphatic leukemia is a leukemic T cell with an altered mechanism of IL-2 production at the level of the Pan T2 binding site.
Reorientation of membrane polypeptides during erythrocyte maturation
Authors: Allen RW , Hoover BA
Published: October 2019

Messenger RNA extracted from the erythroleukemic cell line K562 was translated in vitro and the translation products reacted with an antiserum raised against human erythrocyte ghosts. Polypeptides immunoprecipitated by the antiserum were characterized by SDS- polyacrylamide gel electrophoresis and fluorography. The antiserum immunoprecipitated polypeptides with nominal molecular weights of 37,000 (p37), 20,000 (p20), 19,000 (p19), 18,000 (p18), 14,000 (p14), 13,000 (p13), and 11,000 (p11) daltons. Since the antiserum was raised against antigenic determinants present on both the inner and outer surface of the red cell membrane, differential absorption of antiserum with intact red cells, or ghosts, was used to localize the translation products to the inner or outer membrane surface. Absorption was also used to determine if any of the immunoprecipitated translation products represented membrane markers for the erythroid lineage. Absorption of the antiserum with red cell ghosts removed all antibodies reacting with in vitro translation products. Absorption with intact cells from various lineages removed anti-p20 antibodies and did not absorb anti- p19 or anti-p18 antibodies. Absorption with intact cells from all lineages except mature erythrocytes absorbed anti-p37, anti-p14, and anti-p13 antibodies, suggesting that these antigens are expressed on the outer membrane surface. Mature erythrocytes were incapable of absorbing these antibody populations, suggesting a lineage-specific reorientation of these antigens in the membrane during erythropoiesis.
Surface antigenic determinants on human pluripotent and unipotent hematopoietic progenitor cells
Authors: Bodger MP , Izaguirre CA , Blacklock HA , Hoffbrand AV
Published: October 2019

RFB-1 is a monoclonal antibody previously shown to react with granulocyte-monocyte progenitors (CFU-GM) and immature lymphoid cells in human bone marrow. RFB-HLA-DR is a monoclonal antibody that reacts with HLA-DR (la-like) antigens. The present study shows that the bone marrow subset reactive with both RFB-1 and RFB-HLA-DR contains all the cells that give rise to mixed hematopoietic colonies (derived from CFU- GEMM; a pluripotent human progenitor cell) as well as to megakaryocytic (megakaryocyte-CFU-derived) and erythropoietic (derived from erythroid burst-forming units, BFU-E) colonies, as shown by fluorescence- activated cell sorting and complement-mediated cytotoxicity. These results indicate that CFU-GEMM, BFU-E, and megakaryocyte-CFU express RFB-1 and la-like antigens. RFB-1 antigen is also expressed on erythroid colony-forming units (CFU-E). RFB-1 and RFB-HLA-DR are useful reagents in the study of hematopoietic stem cells.
Derivation of lymphoma cell lines from microscopically normal bone marrow in patients with undifferentiated lymphomas: evidence of occult bone marrow involvement
Authors: Benjamin D , Magrath IT , Douglass EC , Corash LM
Published: October 2019

Evidence is presented that at least 17% of microscopically normal bone marrow samples obtained from patients with undifferentiated lymphomas contain occult tumor cells. Of 19 microscopically normal bone marrow samples tested, continuous tumor cell lines were obtained from 4. A tumor cell origin was confirmed by the presence of an 8;14 chromosomal translocation in each case, and HLA typing confirmed the patient origin of the cell line. In two other patients, direct cytogenetic examination of microscopically normal bone marrow samples revealed karyotypes containing 8:14 translocations or a 14q+ chromosome. These findings indicate that undifferentiated lymphomas are often more widespread than is clinically appreciated. The presence of submicroscopic marrow involvement is also of significance to the design and analysis of treatment protocols involving autologous marrow infusion.
Red blood cell glucose metabolism in trisomy 10p: possible role of hexokinase in the erythrocyte
Authors: Magnani M , Stocchi V , Piatti E , Dacha M , Dallapiccola B , Fornaini G
Published: October 2019

Red blood cell glucose metabolism was investigated in a male patient with de novo trisomy 10p. According to previous evidence, when assigning hexokinase gene locus in the 10p11 leads to pter region, a triplex dosage effect of hexokinase activity (HK) was found, while all the other erythrocyte glycolytic enzymes were in the normal values range. Red blood cell glucose utilization was 2.87 mumole/hr/ml RBC as compared to 1.43 in normal controls; the rate of glucose metabolized through the hexose monophosphate shunt (HMPS) was unchanged. Glucose-6- phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, and dihydroxyacetone phosphate increased with respect to normal controls, while normal levels of 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, and ATP were found. The HK activity increased in all the red blood cell fractions obtained by density gradient ultracentrifugation. However, a small difference in the distribution of cells through the gradient was evident. The experiments reported in this article show that in the red blood cells of patients with trisomy 10p, an increased level of HK leads to higher concentrations of glucose- 6-phosphate and to a faster glucose utilization in the Embden-Meyerhof pathway, while the HMPS rate is unchanged.