bluuebunny/crossref_metadata_2025 huggingface dataset:
Protein C deficiency resulting from possible double heterozygosity and its response to danazol
Authors: Gruppo RA , Leimer P , Francis RB , Marlar RA , Silberstein E
Published: October 2019

A unique family with protein C (PC) deficiency is described. The proband had a history of renal vein thrombosis as a newborn and iliofemoral thrombosis at the age of 6 years. After 6 months of heparin treatment, discontinuation of anticoagulation therapy was accompanied by persistent hypofibrinogenemia with increased fibrinogen consumption. With continuous infusion of heparin, fibrinogen turnover normalized, and the child has remained free of thrombosis. Both the immunologic level of PC and the functional activity measured by amidolytic assay were moderately reduced (47% and 34%, respectively). Functional activity of PC measured by its anticoagulant activity was disproportionately lower (14%). A 3-year-old asymptomatic sibling had a similar disproportionate reduction of PC anticoagulant activity compared with the amidolytic activity or immunologic level. The mother demonstrated type I PC deficiency with a proportionate reduction in immunologic protein levels (59%), anticoagulant activity (52%), and amidolytic activity (46%), whereas the father had type II PC deficiency with normal immunologic protein levels (102%), normal amidolytic function (98%), but a low anticoagulant function (50%). An abnormal PC molecule was detected by two-dimensional immunoelectrophoresis in the father and two children. These data are consistent with the hypothesis that the children are doubly heterozygous for two different types of PC deficiency inherited from each of the parents. A 14-day trial of danazol in the proband resulted in a rise in the PC antigen concentration from 66% to 98% but no change in PC anticoagulant function.
Plasmids in bacteria exposed to activated neutrophils mediate mutagenesis when transferred to new hosts
Authors: De Togni P , Fox HB , Morrissey S , Tansey LR , Levy SB , Babior BM
Published: October 2019

The plasmid pUC18 contains a lacZ alpha-complementation gene that codes for a small peptide that can complement the delta M15 mutation of the Escherichia coli lacZ (beta-galactosidase) gene, converting bacteria carrying that mutated gene from the lacZ- to the lacZ+ phenotype. This plasmid was used in experiments designed to study mutagenesis by human neutrophils. E coli carrying pUC18 were incubated with neutrophils under conditions in which little ingestion of the bacteria took place; the plasmid was then isolated and transformed into an E coli strain (BOZO) that carries the lacZ delta M15 mutation. Of these transformants, 11 of 205,000 were lacZ, suggesting that in these 11, alpha-complementation had been lost through a mutation. No lac- colonies were detected among several hundred thousand BOZO transformed with plasmid isolated from incubations in which phagocytosis could take place, nor from incubations from which neutrophils were omitted. Despite the lac- phenotype of these 11 transformants, plasmids reisolated from nine of them showed normal alpha-complementing ability when transformed into fresh BOZO. These findings indicated that in these nine, the mutations were located in the chromosomes of the transformed BOZO. It thus appears that on exposure to activated neutrophils, a plasmid may acquire a lesion (? mutation) that can somehow be transferred to the genome of a recipient microorganism, resulting in repair of the damaged plasmid accompanied by mutation of the recipient's chromosome. Restriction mapping of the DNA from four of these nine chromosomal mutants suggested that the mutations did not represent major insertions or deletions in the portion of the bacterial chromosome corresponding to the pUC18 lac operon insert, nor in the remainder of the lacZ delta M15 gene. These results confirm previous work showing that exposure to activated neutrophils can induce mutations in biological systems, and provides an experimental model in which the mechanism of neutrophil-mediated mutagenesis may be examined.
Evidence for posttranslational control of Hb C synthesis in an individual with Hb C trait and alpha-thalassemia
Authors: Liebhaber SA , Cash FE , Cornfield DB
Published: October 2019

The level of Hb C in the erythrocytes of individuals with Hb C trait decreases significantly in the presence of coexisting alpha- thalassemia. This relationship may result from the higher affinity of beta A than beta C for limiting amounts of alpha-globin during hemoglobin assembly. This mechanism would predict that the beta A and beta C synthetic capacity in alpha-thalassemic individuals with Hb C trait should be balanced despite the low levels of Hb C in their circulating erythrocytes. To directly test this prediction, we have measured the beta A and beta C synthetic capacity of reticulocyte RNA isolated from two individuals with Hb C trait, one with a normal alpha- globin genotype and one with alpha-thalassemia. The balanced expression of beta A and beta C in both cases supports the proposed posttranslational control over Hb C expression.
Detection of carriers of the autosomal form of chronic granulomatous disease
Authors: Verhoeven AJ , van Schaik ML , Roos D , Weening RS
Published: October 2019

The NADPH:O2 oxidoreductase catalyzing the respiratory burst in activated phagocytes from healthy individuals is not operative in phagocytes from patients with chronic granulomatous disease (CGD). In a microscopic slide test using the dye nitroblue tetrazolium (NBT), carriers of X-linked CGD can be recognized by a mosaic pattern of NBT- positive and NBT-negative cells, governed by the expression of an unaffected or an affected X chromosome, respectively. Until now, it has not been possible to detect carriers of the autosomal form of CGD (other than by family studies) because all cells of these carriers stain positive in the NBT test. We have investigated whether neutrophils from carriers of autosomal CGD can be recognized by measurement of the rate of oxygen uptake upon stimulation of the cells. It was found that with the phorbol ester PMA as a stimulus, the respiratory burst is significantly lower in autosomal CGD carriers. With serum-treated zymosan as a stimulus, no difference between controls and carriers was observed. The addition of f-Met-Leu-Phe (1 microM) to PMA-activated neutrophils of control donors caused a transient increase in oxygen consumption of about 40%. Under these conditions, an increase of more than 100% was observed in neutrophils from carriers of autosomal CGD. These findings provide a simple method for the detection of carriers of the autosomal form of CGD.
Assessment of proliferation during maturation of the B lymphoid lineage in normal human bone marrow
Authors: Hollander Z , Shah VO , Civin CI , Loken MR
Published: October 2019

Different stages of B lymphoid maturation were identified in normal bone marrow using multiple cell surface markers. The proliferation status of each of these maturational stages was determined by simultaneous quantitative DNA analysis on a flow cytometer. The technique used to quantify these parameters preserved the cell surface immunofluorescence, the light scattering properties and the stoichiometric binding to DNA. The proliferating cells were confined to a distinct population of cells expressing CD10. The number of proliferating cells in these populations was relatively constant among 12 separate bone marrow samples. The data suggest that the timing and rate of proliferation of cells within a single lineage may be a preprogrammed aspect of normal maturation.
Low total protein S antigen but high protein S activity due to decreased C4b-binding protein in neonates
Authors: Schwarz HP , Muntean W , Watzke H , Richter B , Griffin JH
Published: October 2019

Protein S, a vitamin K-dependent cofactor for activated protein C, exists in normal adult plasma in a free anticoagulantly active form and in an inactive form complexed to C4b-binding protein. Immunologic and functional levels of protein S and C4b-binding protein in plasma were determined for 20 newborn infants and compared with adult normal pooled plasma. Total protein S antigen levels averaged 23%, similar to other vitamin K-dependent plasma proteins. However, the protein S anticoagulant activity was 74% of that of adult normal plasma. This apparent discrepancy of activity to antigen was shown to be due to low or undetectable levels of C4b-binding protein, which results in the presence of most if not all of protein S in its free and active form. The relatively high level of anticoagulantly active protein S in infants may enhance the potential of the protein C pathway, thereby minimizing risks of venous thrombosis in this group.
Polyspecific natural antibodies and autoantibodies secreted by human lymphocytes immortalized with Epstein-Barr virus
Authors: Seigneurin JM , Guilbert B , Bourgeat MJ , Avrameas S
Published: October 2019

Recent studies have shown that autoreactive B cells and autoantibodies are present in pathological as well as in normal situations. In the present study, we immortalized human B cell lines from normal individuals and from patients with malignant or benign dysglobulinemia with Epstein-Barr virus and examined, after cloning, the autoantibody reactivities of the immunoglobulins secreted by these cells. Forty-two supernatants were analyzed by enzyme-immunoassay on a panel of 13 self and non-self antigens: trinitrobenzenesulfonic acid (TNP), DNA, L- glutamine, L-alanine, L-tyrosine (GAT), actin, myosin, tubulin, albumin, renin, spectrin, transferrin, thyroglobulin, myoglobin, peroxidase, and by immunofluorescence in tissue sections. Fourteen (33%) of the immunoglobulin-secreting cell lines were found to have an autoantibody function; seven secreted IgM, six IgA, and one IgG. The light chains were of the kappa type in 11 cases. The vast majority of these clones reacted with more than five antigens of the panel and all of them reacted with TNP. No correlation was found between a given isotype and an antibody specificity. More than half of these antibodies also reacted with cellular antigens present in tissue sections. None of the four cell lines secreting monoclonal antiviral antibodies reacted with any of the antigens of the panel. The results indicate that immunoglobulins secreted by human monoclonal lymphoid cell lines can have polyspecific autoantibody functions, similar to those found in normal human polyclonal antibodies, in human monoclonal paraproteins and in natural monoclonal antibodies synthesized by murine or rat clones obtained from physiologically normal animals.
Is there a role for selective vasodilation in the management of sickle cell disease?
Authors: Rodgers GP , Roy MS , Noguchi CT , Schechter AN
Published: October 2019

To test the hypothesis that microvascular obstruction to blood flow at the level of the arteriole may be significant in individuals with sickle cell anemia, the ophthalmologic effects of orally administered nifedipine were monitored in 11 steady-state patients. Three patients with evidence of acute peripheral retinal arteriolar occlusion displayed a prompt reperfusion of the involved segment. Two other patients showed fading of retroequatorial red retinal lesions. Color vision performance was improved in six of the nine patients tested. The majority of patients also demonstrated a significant decrease in the amount of blanching of the conjunctiva which reflects improved blood flow to this frequently involved area. Such improvements were not observable in a control group of untreated stable sickle cell subjects. These findings support the hypothesis that inappropriate vasoconstriction or frank vasospasm may be a significant factor in the pathogenesis of the microvascular lesions of sickle cell disease and, further, that selective microvascular entrapment inhibition may offer an additional strategy to the management of this disorder. We believe a larger, placebo-controlled study with nifedipine and similar agents is warranted.
Synergistic effect of granulocyte-macrophage colony-stimulating factor and 1,25-dihydroxyvitamin D3 on the differentiation of the human monocytic cell line U937
Authors: Zuckerman SH , Surprenant YM , Tang J
Published: October 2019

The human monoblastlike cell line U937 can be induced to differentiate by a variety of agents including gamma-interferon, phorbol esters, retinoic acid, and 1,25-dihydroxyvitamin D3 (VD3). Incubation of U937 with 1 to 1,000 units of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) did not induce macrophage differentiation. A synergistic effect on macrophage differentiation was observed, however, when U937 was cocultured with 10(-8) mol/L VD3 plus 50 U/mL GM-CSF. GM-CSF-plus VD3-treated cells demonstrated significant increases in OKM1 antigen expression, increased chemokinesis and chemotaxis, and increased Fc receptor-mediated erythrophagocytosis. Human peripheral blood monocyte cultures also demonstrated increased OKM1 antigen expression and chemotaxis when incubated with 50 to 500 U/mL of GM-CSF for 48 to 72 hours. VD3, however, was not necessary for the increases in effector function observed for GM-CSF-stimulated monocyte cultures. In distinction to the synergistic effect of GM-CSF on VD3-induced differentiation of U937, recombinant human granulocyte colony-stimulating factor (G-CSF) at comparable concentrations had no augmenting effect over that observed for VD3 alone. These results suggest that GM-CSF, in the presence of other physiological stimuli, can induce significant phenotypic changes in GM-CSF-nonresponsive cells of the monocytic lineage and can increase the effector functions of GM- CSF-responsive peripheral blood monocyte cultures.
Proliferation of normal human promyelocytes and myelocytes after a single pulse stimulation by purified GM-CSF or G-CSF
Authors: Begley CG , Nicola NA , Metcalf D
Published: October 2019

Enriched populations of either normal human promyelocytes and myelocytes or blast cells were obtained by fluorescence-activated cell sorting with the monoclonal antibody WEM-G11. These populations were used to study the effect of pulse stimulation by purified recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) or cross-reacting purified murine granulocyte colony-stimulating factor (G- CSF). Maximal clone formation by promyelocytes and myelocytes was observed in 1-mL agar cultures stimulated continuously with 400 units of either CSF and in cultures of cells that were pulse stimulated by 3,200 units (or greater) of either CSF. Pulse stimulation by 800 units of GM-CSF or G-CSF generated 75% clone formation, and pulse stimulation by 200 units CSF gave 50% clone formation. The majority of clones formed by pulse-stimulated cells were only two cells in size; however, some clones were up to 15 cells in size after a single exposure to CSF. Clone formation was not observed in cultures of blast cell populations after a single pulse stimulation with GM-CSF or G-CSF.
Syngeneic leukocytes together with suramin failed to improve immunodeficiency in a case of transfusion-associated AIDS after syngeneic bone marrow transplantation
Authors: Verdonck LF , de Gast GC , Lange JM , Schuurman HJ , Dekker AW , Bast BJ
Published: October 2019

A 22-year-old man who underwent syngeneic bone marrow transplantation (BMT) for acute lymphoblastic leukemia acquired a human immunodeficiency virus (HIV) infection by transfusion of blood products from a donor at risk. The manifestations were acute encephalopathy together with immune thrombocytopenia in the early posttransplant period, and acquired immunodeficiency syndrome (AIDS) developed 20 months after BMT. Because he had a syngeneic donor, the possibility of reconstituting the immune system was investigated by repeated transfer of healthy syngeneic lymphocytes and by combining repeated transfer of syngeneic lymphocytes with the antiviral agent suramin to protect the infused leukocytes from being attacked by HIV. No improvement was observed clinically or in the patient's immune functions by these efforts.
Autologous peripheral hematopoietic stem cell transplantation restores hematopoietic function following marrow ablative therapy
Authors: Kessinger A , Armitage JO , Landmark JD , Smith DM , Weisenburger DD
Published: October 2019

From ten patients with advanced malignant disease involving the bone marrow, autologous hematopoietic stem cells were collected from the peripheral blood during eight four-hour pheresis procedures and cryopreserved. No manipulations to increase the number of stem cells circulating in the blood were used during the collections. Following marrow ablative chemotherapy or chemoradiotherapy, the autologous cells were thawed and infused intravenously (IV). WBCs reappeared in the circulation at a median of eight days (range seven to 11 days) after stem cell infusion. Two patients died early, whereas the other eight reached normal numbers of circulating granulocytes that have persisted for up to greater than 20 months. These eight patients became independent of RBC transfusions (hemoglobin concentration greater than 10 g/dL) at a median of 27 days (range 11 to 58 days) after transplantation. One patient received platelet transfusions for counts less than 50 x 109)/L, one patient developed a clinical picture of idiopathic thrombocytopenic purpura, and six patients maintained a platelet count greater than 20 x 10(9)/L at a median of 23 days (range 14 to 25 days) following stem cell infusion. This technique allows patients ineligible for autologous bone marrow transplantation due to unacceptable anesthetic risks, prior pelvic irradiation, or bone marrow metastases to receive marrow ablative therapy.
Characteristics and expression of binding sites specific for ferritin H- chain on human cell lines
Authors: Fargion S , Arosio P , Fracanzani AL , Cislaghi V , Levi S , Cozzi A , Piperno A , Fiorelli G
Published: October 2019

Purified recombinant human ferritin composed solely of H subunit was radiolabeled and incubated with proerythroleukemic K562 human cells. A specific binding was detected, and it could be displaced only by ferritins, natural or recombinant, containing large proportion of the H subunit. The specific ferritin H-chain binding was saturable, and cells showed 17,000 to 23,000 binding sites per cell. The affinity constant measured at 37 degrees C was of 3 x 10(8) M-1. Treatment with pronase eliminated the specific binding. The binding sites were expressed in a high number during the cellular exponential phase of growth and progressively decreased to disappear when cells reached the plateau phase. Treatment of the cells with desferrioxamine increased recombinant H-ferritin binding, while iron had little effect. K562 cells induced to differentiate by hemin failed to bind ferritin H. Ferritin H-chain binding capacity is present on various cell lines such as HL60, lung cancer, and hepatoma cells. Analysis of the binding sites by western blotting showed a peptide with apparent mol wt of about 100 kd.
Immunocytochemical demonstration of gamma-melanocyte stimulating hormone-like immunoreactivity in human neutrophilic granulocytes
Authors: Vaalasti A , Johansson O
Published: October 2019

Using the indirect immunohistochemical approach the occurrence of gamma- melanocyte stimulating hormone (MSH)-like immunoreactivity in neutrophilic granulocytes is described.
Restoration of adenylate cyclase responsiveness in murine myeloid leukemia permits inhibition of proliferation by hormone. Butyrate augments catalytic activity of adenylate cyclase
Authors: Inhorn L , Fleming JW , Klingberg D , Gabig TG , Boswell HS
Published: October 2019

Mechanisms of leukemic cell clonal dominance may include aberrations of transmembrane signaling. In particular, neoplastic transformation has been associated with reduced capacity for hormone-stimulated adenylate cyclase activity. In the present study, prostaglandin E, a hormonal activator of adenylate cyclase that has antiproliferative activity in myeloid cells, and cholera toxin, an adenylate cyclase agonist that functions at a postreceptor site by activating the adenylate cyclase stimulatory GTP-binding protein (Gs), were studied for antiproliferative activity in two murine myeloid cell lines. FDC-P1, an interleukin 3 (IL 3)-dependent myeloid cell line and a tumorigenic IL 3- independent subline, FI, were resistant to these antiproliferative agents. The in vitro ability of the “differentiation” agent, sodium butyrate, to reverse their resistance to adenylate cyclase agonists was studied. The antiproliferative action of butyrate involved augmentation of transmembrane adenylate cyclase activity. Increased adenylate cyclase catalyst activity was the primary alteration of this transmembrane signaling group leading to the functional inhibitory effects on leukemia cells, although alterations in regulatory G- proteins appear to play a secondary role.
Lack of vimentin occurring during the intrafollicular stages of B cell development characterizes follicular center cell lymphomas
Authors: Moller P , Momburg F , Hofmann WJ , Matthaei-Maurer DU
Published: October 2019

Reactive germinal-center B cells completely lack vimentin. This observation made by using monoclonal vimentin antibody V9 and a sensitive immunoperoxidase technique led to the investigation of vimentin expression in an unselected series of 73 immunohistochemically proven B cell lymphomas. While small lymphocytic lymphomas regularly expressed vimentin, the neoplastic population of mixed, small and large, and large cell lymphomas often contained various proportions of vimentin-negative tumor cells or failed to express it at all. Against the background of the Lukes-Collins classification and the Kiel classification, both defining and subdividing follicular center-cell lymphomas, there were statistically highly significant correlations between the absence of vimentin and the histology, suggesting a germinal center origin. Our data suggest that loss of vimentin expression is indicative of the transient, follicular center-cell stage of normal and malignant B lymphocytic transformation. The complete absence of vimentin within the neoplastic population of all Burkitt's lymphoma cases examined may serve as an argument for its follicular center stage of differentiation, as was asserted by Lukes and Collins.
Spectrin Oran (alpha II/21), a new spectrin variant concerning the alpha II domain and causing severe elliptocytosis in the homozygous state
Authors: Alloisio N , Morle L , Pothier B , Roux AF , Marechal J , Ducluzeau MT , Benhadji- Zouaoui Z , Delaunay J
Published: October 2019

We report on spectrin Oran (alpha II/21), a new spectrin variant found in an Algerian family. It was characterized by the absence of the spots that classically correspond to the alpha II domain using two- dimensional analysis of spectrin limit digests. On the contrary, the abnormal domain was represented by a new set of spots in the 21-Kd and 16-Kd regions, as demonstrated by Western blots using anti-alpha II domain polyclonal antibodies. Spectrin Oran (alpha II/21) was found in the homozygous state in two children belonging to two separate branches of the family. It yields a severe elliptocytosis. Spectrin self- association was altered. The variant was much more difficult to prove in the heterozygous state, in which it results in no clinical and virtually no morphological symptom. In all four parents involved, however, electrophoretic analysis and Western blots showed the existence of the alpha II 21-Kd and 16-Kd peptides. In one parent, who combines spectrin Oran (alpha II/21) and the alpha II type-2 polymorphism, the two-dimensional spots (52, 39, 34, and 29 Kd) were quantified and appeared reduced by 30%: there was an intermediary decrease of spectrin self-association in this person. In the three other parents, spectrin Oran combined with the alpha II type-1 polymorphism. The alpha II type-1 spots (46, 35, 30, and 25 Kd) appeared in normal range, and spectrin self-association was normal. Along with previous observations, the present data emphasize the large fluctuations of the alpha-variant percentage. Provided spectrin Oran was present in a sufficient proportion, we found an associated alteration of the beta II domain (that faces the alpha II domain in the spectrin dimer): the beta II 65-Kd fragment was reduced and the beta II 52-Kd fragment was reciprocally increased.
Idiotype as a tumor-specific marker in childhood B cell acute lymphoblastic leukemia
Authors: Carroll WL , Link MP , Cleary ML , Bologna S , Carswell C , Amylon MD , Smith SD , Levy R
Published: October 2019

Immunoglobulin (Ig) or idiotype (Id) is a tumor-specific target in those B cell malignancies that express this molecule on their surface. We explored the biology of B cell acute lymphoblastic leukemia (B cell ALL) using Id as a tumor marker. In this report we describe the development of anti-Id monoclonal antibodies (MAB) for two children with B cell ALL. These reagents were used retrospectively to study tumor kinetics and to detect residual disease after chemotherapy. In both cases serum Id values were strikingly high at diagnosis (1.2 mg/mL and 10.8 mg/mL), suggesting that the tumor cells were relatively mature B cells capable of significant antibody production. In both patients the serum Id levels fell with the institution of therapy and confirmed that the patients were in remission. Increasing serum Id predicted relapse four months before conventional methods in patient 1, and Id proved to be a more sensitive measure of tumor burden than Southern blot analysis of rearranged Ig genes in bone marrow samples. Surprisingly, low levels of Id were redetected in the second patient just before completing therapy and have persisted for over a year despite the absence of clinical evidence of recurrent disease. Thus, serum Id levels reflect tumor burden during initial therapy but may not necessarily predict tumor progression after a complete clinical remission.
The population of paroxysmal nocturnal hemoglobinuria neutrophils deficient in decay-accelerating factor is also deficient in alkaline phosphatase
Authors: Burroughs SF , Devine DV , Browne G , Kaplan ME
Published: October 2019

In patients with paroxysmal nocturnal hemoglobinuria (PNH) the RBCs, neutrophils (PMNs), monocytes, and platelets derived from the abnormal clone are deficient in the complement-regulatory protein decay- accelerating factor (DAF). RBC acetylcholinesterase (AChE) and leukocyte alkaline phosphatase (LAP) activities are also characteristically low. DAF, AChE, and LAP are known to be anchored within cell membranes to glycophospholipid-containing phosphatidylinositol (PI).Because PNH progenitors contain DAF that appears to be lost with maturation, it has been proposed that this disorder results from abnormal tethering of these and possibly other proteins to membrane PI. We were puzzled, therefore, that our two PNH patients consistently had normal LAP levels. Consequently, we studied their isolated PMNs to compare DAF and LAP activities in individual cells.PMNs were separated by flow cytometry into DAF-positive and - negative populations by using rabbit anti-DAF antiserum and fluorescein- conjugated goat antirabbit IgG. In both patients the majority of PMNs were DAF deficient, and these cells contained very little alkaline phosphatase activity. In contrast, the smaller, DAF-positive cell populations were phosphatase replete. This is the first demonstration that abnormalities in DAF and LAP activity occur in the same PNH PMN population and strengthens the hypothesis that defective anchoring of proteins to membrane glycophospholipid underlies the pathophysiology of this disorder.
Functional dendritic cells are required for transfusion-induced sensitization in canine marrow graft recipients
Authors: Deeg HJ , Aprile J , Storb R , Graham TC , Hackman R , Appelbaum FR , Schuening F
Published: October 2019

Untransfused dogs given 9.2 Gy total-body irradiation and hematopoietic grafts from DLA-identical littermate donors uniformly achieve sustained engraftment, whereas dogs given three transfusions (Tx) of whole blood from the intended marrow donor 24, 17 and 10 days pretransplant uniformly reject their graft. Sensitization appears to be mediated by peripheral blood mononuclear cells and can be prevented by prior irradiation of the Tx product with UV light, known to inactivate leukocytes, in particular, cells with accessory function. In the present study we investigated which leukocyte population was responsible for Tx-induced sensitization and subsequent marrow graft rejection. Surprisingly, neither monocytes nor macrophages or dendritic cells induced sensitization, and all dogs so treated achieved engraftment; however, all four evaluable dogs transfused with UV- exposed blood to which small numbers of normal dendritic cells (12.5 x 10(3)/kg) were added rejected their marrow graft. Among five dogs given UV-exposed blood and normal monocytes (12.5 x 10(3)/kg) only one rejected its graft, and four achieved sustained engraftment. We conclude that donor dendritic cells are necessary, albeit not sufficient for in vivo sensitization. Sensitization is prevented by elimination or inactivation of dendritic cell.
Human immunodeficiency virus infection in patients with leukemia
Authors: Minamoto GY , Scheinberg DA , Dietz K , Gold JW , Chein N , Gee T , Reich LM , Hoffer J , Mayer K , Armstrong D
Published: October 2019

Eighteen human immunodeficiency virus (HIV)-seropositive patients were found among 211 previously treated adult patients with a variety of leukemias who had been multiply transfused before April 1985. Patients known to be homosexual or intravenous drug users were excluded from this study. The spouse of one HIV-seropositive patient became HIV infected and subsequently developed the acquired immune deficiency syndrome. Patients with leukemia who were multiply transfused before the availability of screening of blood products for HIV antibody should be counseled regarding their individual risks of HIV infection and the risk to sexual contacts.
A transient increase in histone H2A ubiquitination is coincident with the onset of erythroleukemic cell differentiation
Authors: Hensold JO , Swerdlow PS , Housman DE
Published: October 2019

Murine erythroleukemia cells are useful for studying the regulation of erythroid differentiation since these malignant pronormoblasts differentiate to orthochromatic normoblasts when treated with a variety of inducing agents. Changes in chromatin proteins have been described following inducer exposure. The significance of these changes, which are greatest in terminally differentiated cells remains unknown. Ubiquitin is a highly conserved 8.5 kilodalton peptide that is covalently linked to up to 10% of histone H2A. We demonstrate that following exposure of MEL cells to inducers of differentiation, a transient increase in ubiquitination of H2A occurs. This change is coincident with the onset of differentiation. This result suggests that ubiquitination of H2A may have a role in the nuclear changes necessary for erythroleukemic cell differentiation.
Low doses v conventional doses of corticoids in immune thrombocytopenic purpura (ITP): results of a randomized clinical trial in 160 children, 223 adults
Authors: Bellucci S , Charpak Y , Chastang C , Tobelem G
Published: October 2019

A randomized clinical trial was performed in 160 children and 223 adults with acute immune thrombocytopenic purpura (ITP). The role of corticoids at this phase of the disease is still controversial. Therefore, patients were randomized to receive either conventional doses (1 mg/kg/day) of corticotherapy or low doses (0.25 mg/kg/day) for 3 weeks. The remission, defined by platelet count superior to 100,000/microliter was studied for adults and children after 6 months and 12 months, respectively. The statistical analysis showed no significant difference between the two corticotherapy regimens neither in children nor in adults. Overall, 74% of children and 41% of adults were in remission. For the first time in acute ITP, a randomized prospective trial showed that both in children and adults low dose corticotherapy (0.25 mg/kg/day) proves to have the same efficacy as conventional doses (1 mg/kg/day).
Immunocytochemical localization of fibrinogen on washed human platelets. Lack of requirement for fibrinogen during adenosine diphosphate-induced responses and enhanced fibrinogen binding in a medium with low calcium levels
Authors: Suzuki H , Kinlough-Rathbone RL , Packham MA , Tanoue K , Yamazaki H , Mustard JF
Published: October 2019

The association of fibrinogen with washed human platelets was examined by immunocytochemistry during aggregation induced by adenosine diphosphate (ADP) and during deaggregation. The platelets were suspended either in a medium containing 2 mmol/L Ca2+ or in a medium containing no added Ca2+ (20 mumol/L Ca2+). Platelets were fixed at several times during aggregation and deaggregation, embedded in Lowicryl K4M, sectioned, incubated with goat antihuman fibrinogen, washed, reacted with gold-labeled antigoat IgG, and prepared for electron microscopy. To determine whether the method detected fibrinogen associated with the platelets, the platelets were pretreated with chymotrypsin (10 U/mL) and aggregated by fibrinogen; gold particles were apparent not only in the alpha granules but on the platelet surface and between adherent platelets as well. In the medium with 2 mmol/L Ca2+, ADP caused extensive aggregation of normal platelets in the presence of fibrinogen (0.4 mg/mL), and gold particles were evident between the adherent platelets and on the platelet surface; when the platelets deaggregated, gold was no longer present on the surface. In a medium without added Ca2+, ADP caused extensive aggregation in the presence of fibrinogen, and large numbers of gold particles were on the platelet surface and even more between adherent platelets. In this medium, the platelets did not deaggregate, and by five minutes, the granules appeared to be swollen or fused. In the absence of external fibrinogen, ADP caused the formation of small aggregates, and fibrinogen was not detected between adherent platelets. Thus, the association of fibrinogen with the platelet surface enhances platelet aggregation but is not essential for the ADP-induced formation of small aggregates. The association of fibrinogen with platelets is greater under conditions in which platelets release their granule contents and do not deaggregate because both endogenous and exogenous fibrinogen take part in aggregation.
A new procoagulant in acute leukemia
Authors: Falanga A , Alessio MG , Donati MB , Barbui T
Published: October 2019

To verify whether cancer procoagulant (CP), a cysteine proteinase procoagulant distinct from tissue factor (TF), is associated with leukemic cells, we assayed the procoagulant activity of blast cell extracts from 26 patients with different cytological subtypes of acute nonlymphoid leukemia (ANLL) according to the French-American-British classification. All the samples except two shortened the recalcification time of normal human plasma, the effect being significantly greater in the M3 subgroup. The two criteria used to distinguish between CP and TF, independence from factor VII in initiating blood coagulation and sensitivity to cysteine-proteinase inhibitors, were positive in 19 samples from M1, M2, M3, and M4 cytological subtypes. None of the M5 samples fulfilled these criteria. In addition, M1, M2, M3, and M4 samples immunoreacted with an anti-CP goat polyclonal antibody on an Ouchterlony immunodiffusion plate. This study provides the first evidence for a procoagulant other than TF that is associated with leukemic cells.
Excess heme in sickle erythrocyte inside-out membranes: possible role in thiol oxidation
Authors: Kuross SA , Rank BH , Hebbel RP
Published: October 2019

It has been suggested that the development of sickle RBC membrane defects might be related to abnormal amounts of membrane-associated heme (a term we use in its generic sense to include hemoglobins, hemichromes, and free heme). Techniques previously used to measure membrane heme, however, would not distinguish between what is truly membrane-associated and what is merely trapped in RBC ghost preparations. Consequently, we have examined extensively washed inside- out membranes (IOM) prepared from normal and sickle RBC. Approximately 25% of the sickle ghost heme is lost upon conversion to IOM, but sickle IOM still have a significant excess (1.6 +/- 0.3 nmol heme/mg membrane protein compared with 0.7 +/- 0.2 nmol/mg for normal IOM, P less than .001). Amounts of ghost heme are only poorly predictive of amounts of IOM heme (r = .664). Preparation of IOM by using isotonic lysis with saponin yields virtually identical amounts of IOM heme. Small amounts of heme (less than 15%) can be displaced from IOM by using manipulations that elute spectrin, displace electrostatically bound proteins, or cleave the cytoplasmic portion of band 3. Treatment of IOM with dithiothreitol (DTT), however, displaces the most heme (35%), and this is almost reproduced (25% displacement) by the treatment of intact RBC with DTT before IOM preparation. Sequential treatment with all manipulations still leaves about 40% of the heme in sickle IOM, which indicates a compartment more intimately associated with the membrane. At least part of this is free heme without globin, as evidenced by abnormal binding of radiochloroquine to sickle IOM. Conversely, some IOM-associated globin is globin without heme because the measurement of globin per se markedly overpredicts amount of IOM heme. There is a strong correlation between RBC density and amounts of either ghost or IOM heme. Finally, the amount of membrane thiol oxidation (as measured by thiol-disulfide-exchange chromatography) does not correlate at all with ghost heme (r = .105), but it correlates well with IOM heme (r = .877, P less than .001). These data demonstrate that there are abnormal amounts of heme truly associated with sickle RBC membranes, and they are consistent with the hypothesis that this membrane-associated heme participates in the pathobiology of the sickle RBC membrane, particularly those aspects perhaps related to thiol oxidation.
Thrombasthenia with an abnormal platelet membrane glycoprotein IIb of different molecular weight
Authors: Jung SM , Yoshida N , Aoki N , Tanoue K , Yamazaki H , Moroi M
Published: October 2019

We describe an individual with abnormal platelet glycoprotein (GP) IIb of different molecular weight (mol wt), a defect that distinguishes this patient from previously reported thrombasthenics. The patient, a 21-year-old female, has a mild bleeding tendency; her platelets lack adenosine diphosphate (ADP) aggregation and have severely suppressed collagen aggregation but a normal response to ristocetin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of her platelets indicates that they contain two types of GPIIb molecules: one with an abnormal mol wt (122 kd, unreduced; 128 kd, reduced) and one with a normal mol wt (128 kd, unreduced; 118 kd, reduced). Relative to the amount of GPIIb in normal platelets, her platelets contain approximately 35% abnormal GPIIb and 20% normal GPIIb. Fibrinogen binding assays on the patient's platelets indicated that they contained 25% of the normal amount of fibrinogen receptors. Crossed immunoelectrophoresis of the patient's platelets demonstrated the formation of a GPIIb/IIIa complex that was mainly composed of normal mol wt GPIIb and GPIIIa. The patient's father has decreased ADP aggregability, and his platelets also contained both abnormal and normal GPIIb (about 50% of the normal level and about 50% of the normal number of fibrinogen receptors); her mother has only normal GPIIb. These results indicate that the patient has heterozygous GPIIb molecules with an abnormality of GPIIb at the molecular level. Studies on this abnormal GPIIb should provide information about the function of GPIIb and the mechanism of its biosynthesis.
The human genes for GM-CSF and IL 3 are closely linked in tandem on chromosome 5
Authors: Yang YC , Kovacic S , Kriz R , Wolf S , Clark SC , Wellems TE , Nienhuis A , Epstein N
Published: October 2019

As demonstrated by long-range mapping of restriction endonuclease recognition sequences and genomic cloning, we found that the human genes encoding interleukin 3 (IL 3) and granulocyte/macrophage colony- stimulating factor (GM-CSF) are tandemly arrayed on the long arm of chromosome 5, separated by 9 kilobases (kb) of DNA. This close physical linkage of genes with similar structure and biologic function suggests that these cytokines may have evolved from a common ancestral gene. This linkage in evolution of two relatively divergent genes further implies that some of the other lymphokine and cytokine genes that appear to share as much or more sequence similarity than do IL 3 and GM- CSF may be distantly related members of a cytokine gene family.
Use of a monoclonal antibody (GA3) to demonstrate lineage restricted O- glycosylation on leukosialin during terminal erythroid differentiation
Authors: Bettaieb A , Farace F , Mitjavila MT , Mishal Z , Dokhelar MC , Tursz T , Breton- Gorius J , Vainchenker W , Kieffer N
Published: October 2019

A murine monoclonal antibody (GA3) obtained by immunizing mice with cells of the human erythroleukemic cell line K562 is shown to define a 105 kilodalton (kd) membrane antigen on K562 cells that is restricted within the hematopoietic system to the erythroid lineage and to a minor population of CD3, CD4 positive T lymphocytes. Cocapping studies and immunoprecipitation experiments performed with GA3 and L10, an anti- sialophorin monoclonal antibody reacting with leukosialin (Gp 105) on K562 cells, demonstrate that the antigen detected by GA3 on K562 cells is identical to leukosialin. Neuraminidase treatment but not tunicamycin treatment of K562 cells abolishes the expression of the GA3- epitope without affecting the L10-epitope thus providing evidence that terminal sialic acid present on O-linked oligosaccharide chains on Gp 105 is essential for the expression of the GA3-epitope. Further analysis by flow cytometry and immune panning experiments performed on bone marrow cells with GA3 or L10 demonstrate that, in contrast to L10, which reacts with all types of hematopoietic progenitors, the epitope recognized by GA3 is restricted to the erythroid lineage, and appears during erythroid differentiation before glycophorin A on the earliest morphologically recognizable erythroid precursor, the proerythroblast. Our results therefore suggest that O-linked oligosaccharides on leukosialin express lineage restricted and even maturation restricted antigenic structures that might serve as cell lineage specific markers.
Bone marrow transplantation for the treatment of acute nonlymphoblastic leukemia in children aged less than 2 years
Authors: Johnson FL , Sanders JE , Ruggiero M , Chard RL Jr , Thomas ED
Published: October 2019

Eleven children with acute nonlymphoblastic leukemia in first remission who were less than 2 years of age at diagnosis were treated with 120 mg/kg of cyclophosphamide, 12-Gy fractionated total-body irradiation, and marrow transplantation. Seven patients remain in complete remission from 3.5 to 13.8 years posttransplant, four for more than 6.75 years. The immediate posttransplant course was relatively uncomplicated in surviving patients. No child developed severe graft-v-host disease. The major long-term side effect has been a slowing in growth. Although the prognosis for such children with conventional chemotherapy remains poor, intensive cytotoxic therapy and marrow transplantation offers an alternative therapy with a chance for cure.
Identification and characterization of a differentiation antigen in human neutrophils and monocytes
Authors: Nozawa R , Kato H , Ito T , Yokota T
Published: October 2019

Human promyelocytic leukemia (HL-60) cells were induced to differentiate into macrophage-like cells by treatment with 10(-7) mol/L 1,25-dihydroxyvitamin D3 (VD3). A monoclonal antibody (MoAb, 60B8), reactive with the particulate of the differentiated cells but not of the untreated cells, was isolated. The antigen recognized by the MoAb became apparent two days after VD3 treatment, and its concentration increased and peaked on day 6. Human neutrophils, followed by monocytes and differentiated HL-60 cells, showed the greatest abundance of the antigen. Monocytes cultured for eight days in vitro lost the antigen. No 60B8 antigen was seen in other blood cells. The MoAb precipitated two polypeptides with an apparent molecular weight (mol wt) of 15,000 (15 k) and 13 k in the detergent-solubilized, 35S-methionine-labeled lysate of the differentiated HL-60 cells. Double-sandwich type enzyme- linked immunosorbent assay (ELISA) devised for the quantitative assay of 60B8 antigen indicated that some 2% to 5% of neutrophil protein was 60B8 antigen. This antigen was not exposed on the neutrophil cell surface, since the cells were not stained immunofluorescently with either mono- or polyclonal antibody, unless they had become permeable. The neutrophil membrane and the granules were separated on the Percoll density gradient, and the antigen was found localized in the plasma membrane-rich fraction. These findings suggested that 60B8 antigen is a novel differentiation antigen for phagocytic cells.
Recombinant alpha 2-interferon in the treatment of B chronic lymphocytic leukemia in early stages
Authors: Rozman C , Montserrat E , Vinolas N , Urbano-Ispizua A , Ribera JM , Gallart T , Compernolle C
Published: October 2019

Ten previously untreated patients with early B cell chronic lymphocytic leukemia (B-CLL) (seven in Rai's stage 0, three in stage I) were given recombinant alpha 2-interferon (alpha 2IF) (2 X 10(6) U/m2 intramuscularly three times a week for a minimum of 14 weeks) to assess its effectiveness. All patients were evaluable for response to therapy and toxicity. No complete response was achieved. In all cases a definite, although transient reduction in the absolute number of peripheral blood lymphocytes was observed. In eight patients an increase in the absolute number of granulocytes was detected. None of the patients experienced severe hematologic toxicity. Fatigue, malaise, and fever were the more common side effects, but all patients were able to finish their treatment as planned. The results of this pilot study suggest that low doses of recombinant alpha 2-IF have some activity in early and previously untreated B-CLL and that further studies of IF effectiveness in B-CLL seem warranted.
Clonal rearrangement of the T cell receptor beta gene in the circulating lymphocytes of erythrodermic follicular mucinosis
Authors: LeBoit PE , Abel EA , Cleary ML , Hoppe RT , Williams ML , Wood GS , Parslow TG
Published: October 2019

Follicular mucinosis is a condition characterized by the abnormal accumulation of acidic mucopolysaccharides in hair follicles. It is classically described as occurring idiopathically in young persons and within the infiltrates of mycosis fungoides in older individuals. We report a 12-year-old girl who had erythrodermic follicular mucinosis, hypereosinophilia, circulating Sezary cells, and both immunophenotypic and genotypic evidence of T cell neoplasia. Erythrodermic follicular mucinosis may represent an unusual variant of the Sezary syndrome, which to date has not been described in children or adolescents.