bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of neutrophil subpopulations with monoclonal antibodies
Authors: Clement LT , Lehmeyer JE , Gartland GL
Published: October 2019

Two monoclonal antibodies have been produced by the hybridoma technique that recognize subpopulations of human neutrophils. The antibodies, termed 1B5 and 4D1, react with a mean percentage of 57% and 51% of peripheral blood granulocytes, respectively. The antigens recognized appear to be neutrophil specific in that these antibodies do not react with eosinophils, platelets, erythrocytes, monocytes, or nonadherent peripheral blood mononuclear cells. Although the neutrophil subpopulations recognized by these antibodies are nearly identical (coinclusive), the antigenic determinants recognized appear to be different. These monoclonal antibodies to neutrophil subpopulations may prove useful to studying functional heterogeneity among neutrophils as well as for investigations of normal and abnormal myeloid differentiation.
Transplantation for severe combined immunodeficiency with HLA-A,B,D,DR incompatible parental marrow cells fractionated by soybean agglutinin and sheep red blood cells
Authors: Reisner Y , Kapoor N , Kirkpatrick D , Pollack MS , Cunningham-Rundles S , Dupont B , Hodes MZ , Good RA , O'Reilly RJ
Published: October 2019

Three patients with severe combined immunodeficiency (SCID) received transplants of HLA haplotype-mismatched parental bone marrow depleted of T lymphocytes by differential agglutination with soybean agglutinin (SBA) and subsequent E-rosette depletion. Two patients achieved durable engraftment with reconstitution of both humoral and cell-mediated immunity. Neither of these patients developed graft versus host disease (GVHD). The third patient achieved only a transient engraftment with concomitant development of mitogen-responsive lymphocytes of paternal origin. Our experience indicates that depletion of T lymphocytes by this technique can abrogate the potential of histoincompatible marrow grafts to induce lethal GVHD without limiting immunologic reconstitution. It also provides further evidence of nonimmune mechanisms of graft resistance that may necessitate preparative treatment of patients with SCID before transplantation with HLA- mismatched marrow cells.
Lipid accumulation and production of colony-stimulating activity by the 266AD cell line derived from mouse bone marrow
Authors: Hines DL
Published: October 2019

The availability of cloned lines of bone marrow stromal cells could facilitate the analysis of their role in hemopoietic cell development. The 266AD cell line was isolated from a colony of lipid-accumulating bone marrow cells growing in a collagen gel. 266AD cells have subsequently been maintained by passage in tissue culture plastic flasks about every 10 days for greater than 10 mo. Subconfluent cultures of cells are fibroblast-appearing, but in confluent cell sheets, prominent foci of lipid-containing cells develop in both uncloned and four separate cloned cell lines. Supernatants from confluent cultures containing lipid-laden cells contain granulocyte- macrophage colony-stimulating activity (GM-CSA) for normal bone marrow cells and can induce differentiation of Abelson virus transformed murine promonocytic leukemia cells. 266AD cells were originally isolated in the presence of hydrocortisone, but hydrocortisone is not necessary for lipogenesis to occur. Growth of bone marrow cells in a collagen gel matrix provided a way to isolate stromal cells, and the 266AD cell line provides a means to examine the relationships between stromal cell lipogenesis and regulation of granulopoiesis.
Chronic granulomatous disease due to granulocytes with abnormal NADPH oxidase activity and deficient cytochrome-b
Authors: Seger RA , Tiefenauer L , Matsunaga T , Wildfeuer A , Newburger PE
Published: October 2019

A patient with an X-linked genetic disease resembling chronic granulomatous disease (CGD) but differing in several aspects from previously studied cases is described. The oxidase enzyme of the patient's granulocytes was normally activated, but had reduced activity as shown by an increased Michaelis constant and decreased maximum velocity of NADPH-dependent superoxide production. Cytochrome-b was undetectable in dithionite difference spectra. This CGD-like disease further implicates cytochrome-b as an important component of the microbicidal NADPH oxidase system and provides insight into its role in the enzyme complex.
Plasma and urine cyclic nucleotide levels in patients with acute and chronic leukemia
Authors: Peracchi M , Lombardi L , Maiolo AT , Bamonti-Catena F , Toschi V , Chiorboli O , Mozzana R , Polli EE
Published: October 2019

Plasma and urine levels of cyclic adenosine 3′,5′-monophosphate (cAMP) and of cyclic guanosine 3′,5′-monophosphate (cGMP) were measured in 35 normal subjects, in 24 patients with nonneoplastic diseases (iron deficiency anemia, peptic ulcer, and cholelithiasis), and in 50 leukemic patients. The leukemic group included patients with acute lymphoblastic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, and chronic myelogenous leukemia. All patients were recently diagnosed and untreated, except for 5 patients with blastic transformation of chronic myelogenous leukemia who had been previously treated. There were no significant differences in plasma and urine cyclic nucleotide levels between normal subjects and patients with nonneoplastic diseases. In leukemic patients, plasma and urine cAMP levels were similar to those of normal subjects, whereas plasma and urine cGMP levels were markedly elevated. There were no significant differences in cGMP values between the various types of leukemia. After starting treatment, plasma cyclic nucleotide levels were periodically measured in 21 of the patients with acute leukemia; cGMP levels were normalized in all the 16 subjects who attained complete remission, whereas both cAMP and cGMP levels were apparently unaffected in the patients who did not respond to treatment. This suggests that plasma or urine cGMP could be used as an additional parameter to monitor the patient's response to treatment.
Bone marrow ablation followed by allogeneic marrow grafting during first complete remission of acute nonlymphocytic leukemia
Authors: Forman SJ , Spruce WE , Farbstein MJ , Wolf JL , Scott EP , Nademanee AP , Fahey JL , Hecht T , Zaia JA , Krance RA , Findley DO , Blume KG
Published: October 2019

Of 33 patients who had undergone allogeneic bone marrow transplantation during first complete remission of acute nonlymphocytic leukemia, 21 patients have now been followed in continued complete remission for 6- 64 mo (median greater than 18 mo) without maintenance chemotherapy. The median age of the surviving patients is 27 yr. Transplant-related complications occurring throughout the first year after marrow grafting were fatal in 7 patients, and leukemic recurrence led to the death of 5 patients. The actuarial long-term disease-free survival is 60% and the actuarial remission rate is 79%.
Pre-B-cells in normal human bone marrow and in bone marrow from patients with leukemia in remission: persistent quantitative differences and possible expression of cell surface IgM in vitro
Authors: Pearl ER
Published: October 2019

Pre-B-cells are bone marrow lymphoid cells that lack surface immunoglobulin (sIg-) but contain intracytoplasmic (c) IgM heavy chains and are probably the immediate precursors of immature sIgM+ B lymphocytes. To better understand early stages of B-cell development, immunofluorescence techniques were employed to identify pre-B-cells and B lymphocytes and to examine the expression of sIgM in vitro by human marrow that had been previously depleted of B cells by immunoadsorption. Marrow was derived from patients with acute leukemia in long-term remission off therapy and from a variety of controls. The pre-B-cell compartment was greatly expanded in the marrow of leukemia remission patients for more than 2 yr following cessation of therapy. A similar finding was noted in two patients with lymphoma who had also completed chemotherapy, but not in three with solid tumors prior to therapy. sIgM+ B cells appeared in cultures of sIg- marrow cells from leukemia patients, but not the controls, and only after exposure to Epstein-Barr virus (EBV). At least some of the sIgM+ lymphocytes also expressed cIgM and were probably derived from pre-B-cells. The results of this study (A) confirm that patients who have completed treatment for acute leukemia have a prolonged elevation of pre-B-cell proportions, (B) demonstrate that similar abnormalities may exist in patients with certain solid tumors following chemotherapy, and (C) suggest that a fraction of sIg- human marrow cells, perhaps pre-B- cells, bear a receptor for EBV and can be induced to express to sIgM in vitro.
Increase of Na-K-ATPase activity, glutamate, and aspartate uptake in dog erythrocytes associated with hereditary high accumulation of GSH, glutamate, glutamine, and aspartate
Authors: Maede Y , Inaba M , Taniguchi N
Published: October 2019

We have found convincing evidence for the presence of Na-K-ATPase and high potassium (K) and low sodium (Na) concentrations in the erythrocytes of some dogs associated with hereditary high concentrations of erythrocyte glutathione and some amino acids, glutamate, glutamine, and aspartate. The Na-K-ATPase activity of the erythrocyte membranes of the dogs was about 3 times that of human erythrocyte membranes, whereas the enzyme activity was not detected in control dogs with a normal level of blood glutathione. The Michaelis constant of the enzyme for ATP (Km ATP) was 6.6 X 10(-3)M in the dogs' erythrocytes and 5.0 X 10(-4)M in the human erythrocytes. The concentration of K in the erythrocytes in the dogs examined was about 11 times that of the controls, whereas the erythrocyte Na concentration in the dogs was about one-third that of the controls. The concentrations of K and Na in the plasma of the dogs were equal to those of the controls. Furthermore, L-3H-glutamate and L-3H-aspartate uptake by those cells with high activity of Na-K-ATPase greatly increased, while L-3H-glutamine uptake was unchanged. It appeared that Na+ and K+ gradients created by Na-K-ATPase across the cell membrane might stimulate glutamate and aspartate uptake by the cells, thus causing the high accumulation of such amino acids in the cells.
The ultrastructural localization of immunoglobulin in chronic lymphocytic lymphoma cells: changes in light and heavy chain distribution induced by mitogen stimulation
Authors: Newell DG , Harris AH , Smith JL
Published: October 2019

The distribution of light and heavy immunoglobulin chains in chronic lymphocytic leukemia (CLL) cells has been investigated at the ultrastructural level using an immunoperoxidase technique. Light chains were localized in the lumens of the perinuclear space and rough endoplasmic reticulum, while staining for heavy chains was weak or negative and generally confined to the membranes of the rough endoplasmic reticulum. This pattern is consistent with immunoglobulin biosynthetic studies on CLL cells in which light chains are synthesized in excess over heavy chains and secreted as the exclusive immunoglobulin product. The pokeweed mitogen stimulation of two populations of CLL cells for 6 days resulted in a balanced synthesis and secretion of light and heavy chains that was reflected in concomitant change in light and heavy chain distribution and intensity of staining using the immunoperoxidase technique.
Lack of Rhesus antigen expression by human committed erythroid progenitors
Authors: Rearden A , Chiu P
Published: October 2019

The D antigen of the Rhesus blood group, an erythroid-specific cell surface marker, is expressed by all morphologically recognizable human nucleated red blood cell precursors including, in low density, the pronormoblast. The object of the present study was to determine the expression of the D antigen by committed erythroid progenitors. Under conditions that produced complete inhibition of BFU-E and CFU-E by known cytotoxic antisera, no significant inhibition was produced by anti-D. Use of anti-human IgG (rabbit) to increase sensitivity and trypsinization to reveal cryptic Rh determinants were both without inhibitory effect. Erythroid bursts and colonies grew normally in methylcellulose that contained anti-D. The addition of anti-D to day 7 BFU-E did not inhibit their proliferation to mature bursts at day 14. These results suggest that the D antigen is not expressed by human committed erythroid progenitor cells. The D antigen is therefore an erythroid-specific differentiation marker, rather than an erythroid- lineage-specific antigen. The development of expression of the D antigen during erythropoiesis parallels that of band 3 protein, to which anti-D has been reported to bind. Lack of Rh expression by committed erythroid progenitors is consistent with the rarity of red cell aplasia in Rhesus hemolytic disease of the newborn and in idiopathic and drug-induced autoimmune hemolytic anemia in which the autoantibodies have apparent Rh specificity. These results imply that Rh compatibility is not a contraindication to human bone marrow transplantation.
The effects of dexamethasone and tetradecanoyl phorbol acetate on plasminogen activator release by human acute myeloid leukemia cells
Authors: Wilson EL , Jacobs P , Dowdle EB
Published: October 2019

This investigation was undertaken to examine the extent to which leukemic cell functions are susceptible to regulation in vitro and to investigate their heterogeneity in this regard. Since plasminogen activator release is known to be a modulatable cellular function that can be influenced by antiinflammatory steroids and tetradecanoyl phorbol acetate (TPA), the effect of these two compounds on the secretion of urokinase- or tissue-type enzymes by leukemic cells was studied. The release of both enzyme species could be stimulated or suppressed by these substances by mechanisms that were inhibitable by actinomycin-D and hence required transcription of new mRNA. Plasminogen activator release by cells from 41/45 patients with AML was either stimulated or inhibited by 10(-7) M dexamethasone, implying that most AML cells possess glucocorticoid receptors. In 26/45 cases, the enzyme was inhibited by this steroid to less than 25% of control values. Pronounced inhibition of this degree was not encountered with normal polymorphonuclear leukocytes. Plasminogen activator secretion by AML cells was profoundly inhibited in 20/41 cases by 1 ng/ml TPA and stimulated in 8/41 cases. Leukemic blasts varied considerably in their response to dexamethasone and TPA. Plasminogen activator release should prove a sensitive means of monitoring the responses of AML cells to biologically active compounds.
Monoclonal antiglycophorin as a probe for erythroleukemias
Authors: Greaves MF , Sieff C , Edwards PA
Published: October 2019

A monoclonal antibody (LICR.LON.R10) specific for the major sialoglycoprotein of the erythroid cell membrane, glycophorin A (alpha), has been used to test the possibility that “cryptic” erythroleukemia may be diagnosed as acute lymphoblastic leukemia (ALL) or acute myeloblastic leukemia (AML). In addition to 27 overt erythroleukemias, 724 leukemias, including 329 ALL (103 in relapse), 205 AML, and 109 blast crises of Ph1-positive chronic myeloid leukemia, were analyzed. Twenty cases with a significant proportion of glycophorin-A-positive (gA+) cells were found; 8 of these (5 AML and 3 blast crises of chronic myeloid leukemia, CML) had an obvious erythroid component, but 12 others were diagnosed as AML (2), AMML (1), CML in myeloid blast crisis (4) or megakaryoblastic blast crisis (1), acute megakaryoblastic leukemia (2), or acute lymphoblastic leukemia (2). The latter two patients had no immunologic evidence supporting a diagnosis of ALL and were resistant to chemotherapy. We conclude that AML and ALL only very rarely express gA, and these are probably genuine “cryptic” erythroleukemias. Other gA+ leukemias (megakaryoblastic and CML blast crises) may arise from bi- or pluripotent stem cells and contain distinct and separable blast cell populations.
The binding of bovine factor XII to kaolin
Authors: Kirby EP , McDevitt PJ
Published: October 2019

Purified bovine factor XII was radiolabeled with iodine-125 and its binding to kaolin studied. Binding was rapid and was not readily reversible upon adding unlabeled factor XII. The optimum pH for binding was in the region of pH 5–7. The isoelectric point of factor XII was pH 5.7. High concentrations of urea or increasing the ionic strength of the medium did not inhibit binding. Polyvalent macromolecules, such as Polybrene and polylysine, were effective inhibitors of factor XII binding to kaolin. Polylysine caused the release of factor XII that had bound to the kaolin surface.
Marrow transplantation from HLA-identical siblings for treatment of aplastic anemia: is exposure to marrow donor blood products 24 hours before high-dose cyclophosphamide needed for successful engraftment?
Authors: Storb R , Prentice RL , Banaji M , Witherspoon RP , Sullivan KM , Stewart P , Sanders JE , Mason M , Doney K , Deeg J , Clift RA , Buckner CD , Appelbaum FR , Thomas ED
Published: October 2019

The present study in patients with aplastic anemia was undertaken to determine whether exposure of recipients to donor blood products 24 hr before preparation with cyclophosphamide (1) enhanced the rate of sustained engraftment of marrow from HLA-identical siblings as suggested by animal experiments, (2) increased the rejection rate, in particular in transfused patients who may already have been exposed to donor antigens by blood products, or (3) was of no relevance to the outcome of transplantation of marrow from HLA-identical siblings. One- hundred fifty-five patients were studied, of whom 78 received blood products from the marrow donor 24 hr before cyclophosphamide and 77 did not. A binary logistic regression analysis was applied to the data, simultaneously considering five previously known risk factors for rejection. Results showed that preceding transfusion of donor blood products had neither a significant beneficial nor detrimental effect on the incidence of sustained engraftment.
Comparative evaluation of fifteen anti-sickling agents
Authors: Chang H , Ewert SM , Bookchin RM , Nagel RL
Published: October 2019

Fifteen compounds reported to be inhibitors of gelation or sickling were studied by standard methods. These tests included (1) the determination of the solubility of deoxyhemoglobin S or Csat, (2) evaluation of sickling in whole SS blood at various pO2s, (3) measurement of the oxygen affinity of hemoglobin and blood, and (4) examination of red cell indices and morphology. Among the 4 noncovalent agents tested, butylurea was the most potent inhibitor of gelation and sickling in vitro; however, relatively high concentrations were required compared to the covalent agents. In the latter group, bis-(3,5 dibromosalicyl)-fumarate, nitrogen mustard, and dimethyladipimidate were especially effective inhibitors of gelation and/or sickling. All of these compounds require further development before they can be considered for clinical use.
Serum monoclonal immunoglobulin bands in undifferentiated lymphomas of Burkitt and non-Burkitt types
Authors: Magrath I , Benjamin D , Papadopoulos N
Published: October 2019

Using an improved electroimmunofixation technique that combines the sensitivity of high resolution agarose gel electrophoresis with the specificity of immunoprecipitation, we have demonstrated monoclonal immunoglobulin bands in the serum of patients with undifferentiated lymphomas of Burkitt and non-Burkitt types. Monoclonal bands were detected in the serum of 12 of 21 patients with extensive tumor, and 1 of 10 patients with minimal tumor. All of the bands were identified as IgM of a single light chain class. Such bands were not detected in the serum of patients with lymphoblastic lymphoma (7) or African Burkitt's lymphoma (6). There was disappearance of the bands after therapy and reappearance at relapse. These findings, coupled with previously reported in vitro information, indicate that undifferentiated lymphoma cells secrete immunoglobulin of IgM isotype. Therefore, such monoclonal bands may be of potential value as tumor markers.
Hertwig's anemia: characterization of the stem cell defect
Authors: Barker JE , Bernstein SE
Published: October 2019

Mice homozygous for Hertwig's anemia, an/an, have a mild macrocytic anemia and are refractory to the potentially curative action of exogenously administered erythropoietin (EP). Both red and white blood cell numbers are decreased. One possible explanation is that a precursor stem cell population is in short supply. We have examined this possibility through comparisons of CFU-S, BFU-E, and CFU-E numbers in an/an and +/+ mouse bone marrow. The response of erythroid-committed stem cells to EP in vitro has also been determined. Our findings suggest that EP resistance is due to diminished numbers rather than decreased EP sensitivity of responsive cells. In mice with Hertwig's anemia, the relative numbers of both BFU-E and CFU-E are reduced. The erythroid-committed stem cells are more severely affected than their progenitors, the CFU-S. The data show that cells are lost during the progressive differentiation from stem cell to erythrocyte.
Relationship among plasma iron, plasma iron turnover, and reticuloendothelial iron release
Authors: Uchida T , Akitsuki T , Kimura H , Tanaka T , Matsuda S , Kariyone S
Published: October 2019

A circadian rhythm was demonstrated in 10 males and 10 females with respective mean decreases in plasma iron concentration at 18 hr of 62% and 47% of morning values. Ferrokinetic studies performed on 5 normal males and 5 normal females showed a more rapid disappearance rate and lower plasma iron turnover in the evening. Parallel studies were done on 6 normal males in the morning and 4 normal males in the evening of the release of reticuloendothelial iron at 8 and 18 hr after intravenous injection of 59Fe chondroitin ferrous sulfate. The 6-hr release in the morning was 54.1% and in the evening 25.9%. Composite data from morning and evening showed a correlation between plasma iron level and plasma iron turnover (r = 0.76, p less than 0.001). A similar correlation existed between the plasma iron level and the percent of radioiron released from the reticuloendothelial system (r = 0.67, 0.02 less than p less than 0.05). These data are consistent with a fluctuating iron release from the reticuloendothelial cell in normal subjects, which would account for the diurnal variation in plasma iron.
Thrombin proteolysis of purified factor viii procoagulant protein: correlation of activation with generation of a specific polypeptide
Authors: Fulcher CA , Roberts JR , Zimmerman TS
Published: October 2019

Factor VIII procoagulant protein (VIII:C) purified from commercial factor VIII concentrate contained multiple polypeptides ranging in mol wt from 79,000 to 188,000, all of which were removed from solution by a monoclonal anti-VIII:C antibody specific for a thrombin-sensitive epitope. In a time-course digest of the purified VIII:C using a trace amount of purified human alpha-thrombin, changes occurred in all VIII:C polypeptides during the activation and inactivation of VIII:C activity. The generation and destruction of a mol wt 92,000 polypeptide paralleled the increase and decrease in VIII:C activity, suggesting that this polypeptide represents an activated form. These results provide the basis for a working hypothesis for the mechanism of thrombin activation of VIII:C.
Alpha-1-antitrypsin-human leukocyte elastase complexes in blood: quantification by an enzyme-linked differential antibody immunosorbent assay and comparison with alpha-2-plasmin inhibitor-plasmin complexes
Authors: Brower MS , Harpel PC
Published: October 2019

An enzyme-linked immunosorbent assay (ELISA) has been developed for the quantification of alpha 1-antitrypsin-human leukocyte elastase (alpha 1AT-E) complexes. In the ELISA, the alpha 1AT-E complex is bound to a surface by rabbit antileukocyte elastase antibody, and the inhibitor- proteinase complex is quantified by a second antibody, rabbit anti- alpha 1-antitrypsin F(ab')2, labeled with alkaline phosphatase. alpha 1AT-E complexes were detected when a final concentration of 2.2 nmol/liter of leukocyte elastase was added to plasma. The concentration of these complexes increased with additional elastase. In clotting blood, alpha 1AT-E complexes were generated in parallel with the conversion of 125I-fibrinogen to fibrin, whereas alpha 2-plasmin inhibitor-plasmin (alpha 2PI-P) complexes were not formed. The concentration of alpha 1AT-E complexes in 19 of 21 controls was less than 2.2 nmol/liter. Patients with laboratory evidence for disseminated intravascular coagulation (DIC) demonstrated elevated alpha 2PI-P complexes with either increased or normal concentrations of alpha 1AT-E complexes. Patients without evidence for DIC, but who demonstrated prolonged reptilase clotting times, were studied. This group had increased alpha 1AT-E but normal alpha 2PI-P complex levels, raising the possibility that elastase release in vivo may be accompanied by limited degradation of fibrinogen. These assays thus serve as useful probes for the study of leukocyte activation and of the interactions between cellular and plasma proteolytic enzyme systems.
Protein kinases and membrane protein phosphorylation in normal and abnormal human erythrocytes: variation related to mean cell age
Authors: Fairbanks G , Palek J , Dino JE , Liu PA
Published: October 2019

Protein kinase activities and membrane autophosphorylation reactions of normal and abnormal human erythrocytes were analyzed. Erythrocytes from patients with high reticulocytosis due to sickle cell anemia and other disorders (n = 13) exhibited elevated activities of total and membrane- bound cAMP-independent casein kinase and cAMP-stimulated histone kinase. Relative to normal controls (n = 10), the average total activities in these abnormal cells were increased 50% and 81%, respectively. The casein and histone kinase activities of normal and abnormal erythrocytes declined significantly with increasing age and buoyant density in Stractan density gradients. Casein kinase activity was highly correlated (r = 0.88; n = 23) with the percentage of reticulocytes in the fraction, consistent with either a progressive loss of activity in mature erythrocytes or an abrupt decline during reticulocyte maturation. The cAMP-independent and cAMP-stimulated autophosphorylation activities of isolated membranes also declined with increasing erythrocyte age. On average, the initial rate of spectrin labeling was 36% lower in ghosts from Stractan gradient bottom fractions, relative to ghosts from top fractions similarly incubated with gamma-32P-ATP. Incorporation into the “band 4.5 zone” (primarily labeling bands 4.8 and 4.9, mol wt 47,800 and 44,600) was also age- dependent. In membranes of unfractionated sickle cells, spectrin autophosphorylation was within normal limits, while 4.5 zone autophosphorylation was increased. Membranes from high reticulocytosis controls (vitamin B-12 deficiency) exhibited similar autophosphorylation patterns, suggesting that the altered autophosphorylation pattern of sickle cell membranes may be attributed to the predominance of very young cells.
C-peptide does not parallel increases of serum levels of substances immunologically cross-reactive with insulin in non-Hodgkin's lymphoma patients
Authors: Pavelic K , Vuk-Pavlovic S
Published: October 2019

In 28 of 45 patients suffering from non-Hodgkin's lymphoma, the blood levels of substances detectable by the insulin-specific radioimmunoassay were supranormal, while the C-peptide levels were almost normal or even low. Such a ratio of these substances was also found in a diabetic non-Hodgkin's lymphoma patient. Two sera with the highest concentrations of these substances were investigated by gel filtration. In one case, the insulin cross-reactive material was eluted at the position of the insulin marker; in the other serum, its molecular mass was approximately 120,000.
Allogeneic marrow transplantation for acute nonlymphoblastic leukemia after first relapse
Authors: Appelbaum FR , Clift RA , Buckner CD , Stewart P , Storb R , Sullivan KM , Thomas ED
Published: October 2019

Sixty-two patients with acute nonlymphoblastic leukemia in first relapse or second remission were treated with allogeneic marrow transplantation from HLA-matched siblings. In 17 patients (group 1), no attempt at reinduction of remission was made prior to transplantation. In 20 patients (group 2), attempts at inducing a second remission prior to transplantation were unsuccessful; and in 25 patients (group 3), a second remission was achieved. Five of 17 patients (29%) in group 1, 2 of 20 (10%) in group 2, and 5 of 25 (20%) in group 3 are surviving disease-free 2–6 yr after grafting. Early mortality from nonleukemic causes was equal in the 3 groups, but the risk of recurrent leukemia after transplantation was less in patients transplanted without attempts at reinduction (group 1). Among patients transplanted in relapse, those in early relapse (less than 30% blast cells in the marrow) appeared to do better than patients in florid relapse. The results obtained in group 1 are as good as or better than those achieved in patients transplanted in second or subsequent remission. Thus, for patients with acute nonlymphoblastic leukemia not transplanted in first remission, the optimal time for transplantation would appear to be as soon as possible after the first relapse.
Fanconi's anemia treated by allogeneic marrow transplantation
Authors: Deeg HJ , Storb R , Thomas ED , Appelbaum F , Buckner CD , Clift RA , Doney K , Johnson L , Sanders JE , Stewart P , Sullivan KM , Witherspoon RP
Published: October 2019

Eight patients with Fanconi's anemia were given cyclophosphamide alone (seven patients) or combined with procarbazine and antithymocyte globulin (one patient) followed by marrow grafts from HLA-identical siblings. All patients had engraftment. Seven developed acute and three chronic graft-versus-host disease (GVHD). Three patients died with GVHD and infectious complications (days 19, 56, and 82) and one with an intracerebral hemorrhage (day 540). Four patients are surviving 647- 3435 days after grafting, two are well, and two have chronic GVHD that is improving. These results show that Fanconi's anemia can be treated successfully by allogeneic marrow transplantation.
A technique for specific removal of factor IX alloantibodies from human plasma: partial characterization of the alloantibodies
Authors: Theodorsson B , Hedner U , Nilsson IM , Kisiel W
Published: October 2019

A method for specific removal of large amounts of factor IX:C alloantibodies by a resin to which highly purified factor IX was linked (factor IX CH-Sepharose) is described. Factor IX was isolated from human plasma by a three-step procedure, including barium citrate adsorption and elution, DEAE-Sepharose CL-6B chromatography, and dextran sulfate agarose chromatography. Approximately 100 mg factor IX was obtained from 60 liters of plasma. The preparation was about 95% pure as judged by SDS-PAA gel electrophoresis. Its specific coagulant activity was 160 U/mg (IX) and its factor IX clotting antigen (IX:Ag) 500–600 U/mg. Essentially quantitative coupling of the factor IX preparation to activated CH-Sepharose 4B was obtained (4 mg factor IX/ml gel; 2300–3000 U/IX:Ag/ml). This resin bound 1500–2000 U factor IX inhibitor/ml gel and could be re-used at least 5 times without any loss in binding capacity. The binding capacity was dependent on the flow rate. No signs of activation of the coagulation, fibrinolytic, or complement system were observed in vitro. Using this factor IX resin, factor IX alloantibodies were isolated and found to consist of two portions, one minor bound to the resin only in the presence of Ca2+ and another major portion Ca2+ independent. The specific inhibitory activity/milligram IgG of the Ca2+-dependent alloantibodies was about 5 times higher in the presence of Ca2+. It is concluded that 25 ml of the factor IX resin described can remove about 40,000 factor IX inhibitor units (comparable to 120,000 Bethesda U) in one run, provided the flow rate does not exceed 20 ml/hr. By using such a technique for removal of antibodies it seems feasible to convert hemophilia-B patients complicated with inhibitors against factor IX into ordinary hemophilia- B patients for treatment at an emergency or in association with major surgery.
Selective stimulation by mouse spleen cell conditioned medium of human eosinophil colony formation
Authors: Metcalf D , Cutler RL , Nicola NA
Published: October 2019

Stimulation of unfractionated or nonadherent human marrow cells in agar culture by pokeweed-mitogen-stimulated BALB/c mouse spleen cell conditioned medium (SCM) led, in most cultures, to the exclusive formation of eosinophil colonies. The culture system exhibited linearity of eosinophil colony formation with varying numbers of cells cultured, and the absolute numbers and size of SCM-stimulated eosinophil colonies approximated those in cultures stimulated by human placental conditioned medium. The active factor in SCM for human eosinophil colony formation was not clearly separable from the factors stimulating granulocyte-macrophage and eosinophil colony formation by mouse marrow cells on ammonium sulfate and phenyl boronate chromatography, but was of larger size than the mouse-active factors and separable from them by phenyl sepharose chromatography. This selective culture system for eosinophil colony formation should be of value for studies on human eosinophil progenitor and maturing cell populations in a variety of disease states.
Lineage infidelity in acute leukemia
Authors: Smith LJ , Curtis JE , Messner HA , Senn JS , Furthmayr H , McCulloch EA
Published: October 2019

Blast cells from 20 patients with acute leukemia (13 diagnosed myeloblastic and 7 as lymphoblastic, using the FAB classification) were studied using antibodies to lineage-specific differentiation markers. The phenotypic findings were usually consistent with the clinical diagnosis. However, examples were encountered where individual blast cells had a cytoplasmic marker of one lineage and a surface marker of a different lineage (lineage infidelity). Six examples of intramyeloid (two different myeloid lineages in the same cell) and three examples of interlineage infidelity (myeloid and lymphoid markers in the same blast cell) were encountered. No doubly marked cells were found in control material consisting of normal marrow cells, marrow regenerating after transplantation, or multilineage colonies derived from marrow in culture. A significant trend was observed relating the presence of lineage infidelity and failure of remission-induction. The data are interpreted as support for abnormal gene expression in leukemia.
Characterization of the effects of cultured vascular cells on the activation of blood coagulation
Authors: Rodgers GM , Greenberg CS , Shuman MA
Published: October 2019

The coagulant properties of intact bovine vascular cells (aortic endothelial and smooth muscle cells) and human vascular cells (cutaneous and foreskin microvascular cells, umbilical venous endothelium) grown in vitro were studied. Compared to nonvascular cells (fibroblasts, corneal endothelial cells, fetal lung or intestinal mucosal cells), vascular cells had little procoagulant activity. Radioimmunologic measurement of thrombin in recalcified plasma demonstrated markedly lower concentrations of thrombin in the presence of vascular endothelial and smooth muscle cells compared to corneal endothelial and fetal lung cells. The low thrombin concentrations were not a consequence of thrombin binding to the vascular cells nor were they due to accelerated thrombin inactivation by antithrombin-III or alpha 2-macroglobulin. Neither vascular cells nor the nonvascular cells promoted contact activation of plasma as measured by a sensitive specific assay for kallikrein. Studies with intact cell monolayers and purified factors VIIa and X indicated that while nonvascular cells express tissue factor activity, vascular cells do not exhibit this property. These data suggest that the nonthrombogenic nature of intact vascular cells is due to their failure to initiate contact activation and to express tissue factor activity. In addition, the primary difference in coagulant potential between vascular cells and nonvascular cells is the lack of tissue factor expression by the vascular cells.
The expression and modulation of human myeloid-specific antigens during differentiation of the HL-60 cell line
Authors: Graziano RF , Ball ED , Fanger MW
Published: October 2019

Antigenic changes detected by myeloid-specific monoclonal antibodies on HL-60 cells induced to differentiate by various chemical mediators were investigated using flow cytometry. Antigen levels detected by monocyte- granulocyte-specific monoclonal antibodies AML-2–23, 61D3, and 63D3 increased dramatically after differentiation of HL-60 cells along the granulocytic pathway by the addition of dimethyl formamide (DMF), dimethylsulfoxide (DMSO), or cis-retinoic acid. The expression of these same antigens also increased in conjunction with monocytoid differentiation when HL-60 cells were treated with supernatants from leukocytes stimulated with phytohemagglutinin (PHA-LCM) or with mixed lymphocyte conditioned medium (MLC). In contrast, treatment of HL-60 cells with phorbol 12-myristate 13-acetate (PMA), which also induced differentiation along the monocyte pathway, had no effect on the expression of these monocyte-associated antigens. The expression of antigens on HL-60 cells recognized by the granulocyte-specified monoclonal antibodies PMN 6 and PMN 29 decreased after treatment of HL- 60 cells with PMA, but remained constant after treatment with DMF, DMSO, cis-retinoic acid, PHA-LCM, or MLC. These results suggest that normal myeloid differentiation may be dependent on various signals and that morphological and cell surface marker maturity may, under some conditions, be separable. The utility of the HL-60 cell line as a model of myeloid differentiation and for evaluation of inductive signals is discussed.
Fetal hemoglobin production in cultures of primitive and mature human erythroid progenitors: differentiation affects the quantity of fetal hemoglobin produced per fetal-hemoglobin-containing cell
Authors: Dover GJ , Chan T , Sieber F
Published: October 2019

Single-cell microscopic immunodiffusion assays were used to determine the cellular mechanisms that regulate fetal hemoglobin (HbF) levels in cultures of primitive and late erythroid precursors obtained from human adult bone marrow. Two variables--the percentage of cells containing HbF (F cells) and the picograms (pg) of HbF/F cell--were assayed in cells derived from erythroid colony-forming units (CFU-E) and from erythroid burst-forming units (BFU-E) at 7 and 14 days in culture, respectively. The percentage of F cells among all nucleated cells from CFU-E-derived colonies (29.4% +/- 12.5%, mean +/- SD) was not significantly different (p = 0.2) from the percentage of F cells from BFU-E-derived bursts (37.3% +/- 10.1%). Serial daily assays of all cells in cultures on days 3 through 7 and on day 14 revealed a marked increase in F cells between days 4 and 6 in culture. The average amount of HbF/F cell was less in CFU-E-derived F cells than in BFU-E-derived cells (3.5 +/- 0.3 pg versus 6.2 +/- 3.3 pg; p less than 0.01), while adult hemoglobin (HbA) levels in CFU-E-and BFU-E-derived cells remained comparable (19.9 +/- 2.2 pg versus 21.9 +/- 5.3 pg, p = 0.3). These findings indicate that F cell number in culture is not significantly influenced by the relative maturity of the erythroid precursors from which the cells are derived. Differences in the levels of HbF between CFU-E-and BFU-E-derived cells are due to differences in the amount of HbF per F cell, not F cell number.
Lithium enhancement of megakaryocytopoiesis in culture: mediation via accessory marrow cells
Authors: Chatelain C , Burstein SA , Harker LA
Published: October 2019

To examine the effect of lithium (Li) on early megakaryocytopoiesis, murine marrow megakaryocytic (CFU-M) and granulocyte-macrophage (CFU-C) progenitors were assayed in vitro with and without addition of lithium chloride (LiCl) to culture. At 2 mM LiCl, the numbers of CFU-M- and CFU- C-derived colonies were increased to 146% +/- 8% and 128% +/- 6% of controls, respectively (p less than 0.005). Enumeration of megakaryocytes per colony showed a 78% increase of colonies (p less than 0.05) containing from 6 to 22 cells, suggesting an increased proliferative capacity of CFU-M in the presence of LiCl. Conditioned media from spleen cells cultured in the presence of both pokeweed mitogen (PWM-SCM) and 2 mM Li increased the numbers of CFU-M and CFU-C to 157% +/- 8% and 183% +/- 8%, respectively (p less than 0.025), compared to control cultures stimulated by PWM-SCM alone. Since the production of active colony-stimulating activities (CSA) from mitogen- stimulated conditioned media requires T lymphocytes, we hypothesized that the enhancement of the growth of early hematopoietic progenitors in marrow cultures was due to a Li-induced CSA production by accessory marrow cells, rather than a direct effect of Li on stem cells. To test this, cyclosporin-A (CyA), a T-lymphocyte function inhibitor known to suppress CSA production in PWM-SCM, was added to marrow cultures in the presence of 2 mM Li. CyA (3 micrograms/ml) abrogated the Li-induced enhancement of CFU-M and CFU-C growth, but had no effect on colony formation when added alone. The data suggest that the Li-induced enhancement of early megakaryocytopoiesis and granulocytopoiesis is due to local production of CSA(s) by an accessory cell population and requires the integrity of T-lymphocyte function.
The role of ADP secretion and thromboxane synthesis in factor VIII binding to platelets
Authors: Di Minno G , Shapiro SS , Catalano PM , De Marco L , Murphy S
Published: October 2019

Following stimulation with arachidonic acid, collagen, U-46619 (a stable analogue of prostaglandin endoperoxide/thromboxane-A2), thrombin, or adenosine diphosphate (ADP), unstirred human platelet suspensions bound labeled factor VIII in a reaction that reached equilibrium within 10 min. Apyrase inhibited binding induced by arachidonic acid, collagen, U-46619, and thrombin by less than 40%, but inhibited ADP-induced binding by 95%. Binding to aspirin-treated platelets was normal in response to U-46619, reduced by 60%-70% in response to ADP, collagen, and thrombin, and absent in response to arachidonic acid. Binding in response to U-46619 was not altered by the combination of apyrase and aspirin. Binding of factor VIII was decreased by 90% when 10 mM EDTA was added before each agonist, but it was inhibited less than 30% when EDTA was added following platelet stimulation. We conclude that arachidonic acid, collagen, and thrombin can expose binding sites for factor VIII independently of released ADP; that Ca++ is required for activation but probably not for binding of factor VIII to platelets; and that platelet thromboxane synthesis plays a major role in the binding of factor VIII to platelets induced by thrombin, ADP, or collagen.
Inhibition of platelet prothrombinase activity by a lupus anticoagulant
Authors: Dahlback B , Nilsson IM , Frohm B
Published: October 2019

Lupus anticoagulants are spontaneously occurring antibodies with specificity for negatively charged phospholipids. The plasma of a patient with such a polyclonal antibody of IgM type demonstrated low levels of factor VIII coagulant activity (VIII:C) and factors IX, XI and XII when analyzed by biologic clotting assays, whereas in immunochemical assays, normal levels of VIII coagulant antigen and factor IX were obtained. After immunoadsorption of patient plasma with anti-IgM Sepharose, normal biologic activities were demonstrated in clotting assays for VIII:C, factors IX, XI, and XII. The addition of the patient's isolated IgM to normal plasma resulted in grossly abnormal results in these coagulation assays, and a pattern similar to that of the patient's plasma was obtained. The inhibitory effect of the patient's lupus anticoagulant on blood coagulation was demonstrated also in platelet-rich plasma. The results of the clotting assays indicated that the anticoagulant inhibited several of the reactions in the blood coagulation cascade. The availability of purified components made it possible to demonstrate an inhibiting effect on the activation of prothrombin by factor Xa in the presence of isolated platelets, as well as in a system where purified factor V and well defined phospholipid vesicles were substituted for the platelets.
Immunoglobulin phenotype on B cells correlates with clinical stage of chronic lymphocytic leukemia
Authors: Ligler FS , Kettman JR , Smith RG , Frenkel EP
Published: October 2019

The present study examines the relative amounts of surface immunoglobulin (Ig) on lymphocytes obtained from 64 patients with chronic lymphocytic leukemia (CLL) and correlates these findings with the clinical stage of disease. Since the maturing B cell first expresses surface IgM, followed by IgD, and subsequently by IgG, IgA, or IgE, the surface Ig phenotype can be used as a marker of differentiation. Surface Ig was analyzed using the fluorescence- activated cell sorter under carefully controlled conditions. The cells from all patients with CLL were monoclonal with respect to light chain type. Those patients with IgM as the brightest heavy chain class (suggesting relatively immature B cells) had clinically advanced stages of CLL, whereas those with a predominance of IgG (suggesting more mature cells) have a lesser stage of CLL. Thus, the predominant Ig heavy chain class appears to correlate with clinical stage of CLL and provides a clue to the potential aggressiveness of the tumor.
Chemotherapy for acute myelogenous leukemia in children and adults: VAPA update
Authors: Weinstein HJ , Mayer RJ , Rosenthal DS , Coral FS , Camitta BM , Gelber RD
Published: October 2019

We designed a protocol (VAPA) that featured 14 mo of intensive postremission induction chemotherapy in an effort to improve remission durations for patients with acute myelogenous leukemia (AML). One hundred and seven patients under 50 yr of age were entered into this study. The rate of complete remission is 70%. A Kaplan-Meier analysis of patients entering remission predicts that 56% +/- 7% (+/-SE) of patients less than 18 yr and 45% +/- 9% of patients aged 18–50 yr will remain in remission at 3 yr (median follow-up is 43 mo). Patients with the monocytic subtype had a statistically significant shorter duration of remission (2-sided p less than 0.05). There was a high incidence of primary CNS relapse in children. Thirty-one of 41 patients who completed the regimen remain in remission without maintenance therapy. We conclude that the VAPA protocol continues to offer a promising approach to treatment of AML.
Use of an antibody-ricin A-chain conjugate to delete neoplastic B cells from human bone marrow
Authors: Muirhead M , Martin PJ , Torok-Storb B , Uhr JW , Vitetta ES
Published: October 2019

Affinity-purified rabbit antibody to human lambda and kappa chains (R alpha H lambda kappa) was conjugated to the A-chain of the plant toxin, ricin. The resulting immunotoxin (R alpha H lambda kappa-A) killed cells from the tumor cell line Daudi, which bears surface immunoglobulin, but was nontoxic to the CFU-E, BFU-E, and CFU-GM of human bone marrow. R alpha H lambda kappa-A eliminated 99% of clonogenic Daudi cells that had been mixed with marrow cells in vitro, without demonstrable toxicity to hematopoietic cells. Thus, in vitro treatment of marrow with R alpha H lambda kappa-A may increase the incidence of cure following autologous bone marrow transplantation for the treatment of human B-cell malignancies.