bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of monoclonal antibodies to identify cerebrospinal fluid lymphoblasts in children with acute lymphoblastic leukemia
Authors: Homans AC , Forman EN , Barker BE
Published: October 2019

The identification of small numbers of leukemic cells in the cerebrospinal fluid (CSF) presents a diagnostic problem in the treatment of children with acute lymphoblastic leukemia (ALL). We adapted a latex sphere rosetting technique to allow us to identify simultaneously cell surface markers and cell morphology in 199 CSF samples from 34 patients and 14 control subjects. In patients without leukemic meningitis, the majority of CSF lymphocytes (69%) were found to be mature T cells positive for OKT11. A much smaller number of cells (8%) were found to be B cells positive for la. In these children, only 3% of CSF lymphoid cells expressed the common acute lymphoblastic leukemia antigen (CALLA). Similar results were found in the control subjects. By contrast, 28 CSF samples from nine children with varying numbers of CSF lymphoblasts had much greater proportions of CALLA- and la-positive CSF cells (24% to 96%). Leukemic meningitis was present in one of these patients and later developed in four others. However, three patients with small numbers of lymphoblasts present but with low proportions of CALLA-positive CSF cells (less than 5%) subsequently had normal CSF examinations. We found the use of this rosetting technique valuable in providing information complementary to that obtained from cell morphology alone about the possible malignant nature of small numbers of lymphoblast-like CSF cells seen on cytocentrifuge preparations in children with ALL.
Transformed T lymphocytes infected by a novel isolate of human T cell leukemia virus type II
Authors: Chorba TL , Brynes R , Kalyanaraman VS , Telfer M , Ramsey R , Mawle A , Palmer EL , Chen AT , Feorino P , Evatt BL
Published: October 2019

Human T cell leukemia virus type II (HTLV-II) has been isolated from a patient (Mo) with features of leukemic reticuloendotheliosis (LRE) and from a patient with acquired immunodeficiency syndrome (AIDS). We have obtained another isolate of HTLV-II from a patient (CM) with severe hemophilia A, pancytopenia, and a 14-year history of staphylococcal and candidal infections but no evidence of T cell leukemia/lymphoma, AIDS, or LRE. Fresh mononuclear cells and cultured lymphocytes from CM express retroviral antigens indistinguishable by molecular criteria from HTLV-IIMo. Leukocyte cultures from CM yield hyperdiploid (48,XY, +2, +19) continuous lymphoid lines; human fetal cord blood lymphocytes (CBL) are transformed by cocultivation with these CM cell cultures but retain normal cytogenetic constitution. Electron microscopic examination of the CM cultures and transformed CBL reveals budding of extracellular viral particles, intracellular tubuloreticular structures, and viral particles contained within intracellular vesicles. CM cell cultures and the transformed CBL do not require exogenous interleukin 2, have T cell cytochemical features and mature T helper phenotypes, and exhibit minimal T helper and profound T suppressor activity on pokeweed mitogen-stimulated differentiation of normal B cells. These characteristics, which are similar to those observed with the first HTLV-II isolate, may represent properties of all HTLV-II-infected T cells.
Prognostic discrimination among younger patients with chronic granulocytic leukemia: relevance to bone marrow transplantation
Authors: Sokal JE , Baccarani M , Tura S , Fiacchini M , Cervantes F , Rozman C , Gomez GA , Galton DA , Canellos GP , Braun TJ
Published: October 2019

To obtain information relevant to the question of bone marrow transplantation, we examined the prognostic significance of disease features recorded at the time of diagnosis among 625 patients, aged 5 to 45, with Philadelphia chromosome-positive, nonblastic chronic granulocytic leukemia. The actuarial death rate for this population was 5% during the first year after diagnosis, 12% during the second year, and averaged 22.5% per year during the next eight years. Multivariable regression analysis of features recorded in nearly all cases indicated that sex, spleen size, hematocrit, platelet count, and percentage of circulating blasts were significant prognostic indicators. Analyses of additional data available in 113 to 421 cases suggested that serum lactic dehydrogenase activity, percentage of blasts in marrow, nucleated RBCs in blood, and percentage of basophils plus eosinophils might also provide useful prognostic information. A Cox model, generated with five variables representing features recorded regularly (the first five listed), permitted segregation of these patients into three groups with significantly different survival patterns. The high- risk group exhibited an actuarial mortality of 30% during the first two years after diagnosis and an annual risk of 30% thereafter. In contrast, the most favorable group had a two-year actuarial mortality of 9% and an average risk thereafter of 17% per year, with a median survival of 5 1/2 years. We conclude that it should be possible to classify potential candidates for bone marrow transplantation according to risk with conventional therapy. Such information may be useful in making decisions regarding early v deferred marrow transplantation.
Prognostic significance of immunologic phenotype in hairy cell leukemia: does it exist?
Authors: Golomb H , Strehl S , Oleske D , Vardiman J
Published: October 2019

Of 94 hairy cell leukemia (HCL) patients studied for immunologic phenotype of their hairy cells, 89 patients had B cell markers and five patients were both surface immunoglobulin (slg) negative and E rosette negative. Forty of the 77 cases that had the heavy chains individually determined had IgG only (52.0%); 23 others had IgG in addition to other Ig heavy chains. Seventy-nine patients had monoclonal light chains; 65 with kappa chain and 14 with lambda chain. The only significant difference with respect to survival among the various slg groups occurred between the kappa chain and the lambda chain groups. Within the first 46 months after diagnosis of HCL, 20 deaths occurred among the 65 kappa chain patients, whereas the first and only death among the lambda chain patients occurred at 68 months after diagnosis. The only clinical or laboratory parameter that was significantly different between these two immunologic subgroups was the incidence of infections. Among the lambda chain patients, an infectious complication rate of 28.6% was observed subsequent to the diagnosis of HCL, whereas this rate was 68.8% in kappa chain patients (P = .005). The survival of lambda patients was found to exceed that of the kappa patients by the generalized Wilcoxon test (P = .03). However, when the log rank test was used, no significant difference was detected (P = .13).
There are differences in cytogenetic abnormalities among histologic subtypes of the non-Hodgkin's lymphomas
Authors: Levine EG , Arthur DC , Frizzera G , Peterson BA , Hurd DD , Bloomfield CD
Published: October 2019

Although many recurring chromosome abnormalities have been found in malignant lymphoma (ML) in recent years, their relationship to histology remains largely undefined. We have correlated, in the same tumor mass, chromosome findings with histology, defined by the International Working Formulation for Clinical Usage, in 120 patients. We find differences among histologies in the frequency of normal metaphases and the modal number of the predominant abnormal clone. In addition, most histologies have been significantly (P less than .01) associated with specific chromosome abnormalities. In particular, ML, follicular, predominantly small cleaved cell was associated with t(14;18)(q32;q21); ML, follicular, mixed small cleaved cell and large cell with t(14;18)(q32;q21) and trisomy 8; ML, follicular, predominantly large cell with trisomy 7 and breaks in 17q21-q25; ML, diffuse, mixed small cell and large cell with breaks in 11p; ML, diffuse, large cell with trisomy 21 and breaks in 2q and 9q; ML, large cell, immunoblastic with breaks at 6q21; and ML, small noncleaved cell with t(8;14)(q24;q32). Only the associations with t(14;18) and t(8;14) have been previously reported. The associated chromosome abnormality usually occurred in 30% to 70% of a given histology, raising the possibility that cytogenetics may add important prognostic information in lymphoma as it does in the acute leukemias.
The degradation of platelet-activating factor in the plasma of a patient with familial high density lipoprotein deficiency (Tangier disease)
Authors: Pritchard PH , Chonn A , Yeung CC
Published: October 2019

Platelet Activating Factor (PAF) (1-O-alkyl-2-acetyl sn-glycerol 3- phosphocholine) has been characterized by its ability to aggregate platelets at low concentrations and its profound hypotensive effects. There is evidence that the rate of catabolism of this compound in the plasma regulates its concentration. In humans, we and others have shown that a PAF acetylhydrolase is associated with low density lipoprotein (LDL). The LDL particle in the plasma of patients with Tangier disease is quite different from normal as its lipid core appears to be enriched with triacylglycerol. Thus, we have studied the potential of this abnormal lipoprotein to degrade PAF. The assay for PAF acetylhydrolase was based on the release of 3H from PAF that was labelled in the acetate moiety of the sn-2 position. Tangier disease plasma had approximately 3.3-fold higher PAF acetylhydrolase activity (208 +/- 9 nmol/min/mL) than controls (63 +/- 18 nmol/min/mL). This increase was brought about by an increase in the Vmax (400 +/- 40, Tangier disease; 54 +/- 5, controls) and Km for PAF (120 +/- 20 mumol/L, Tangier disease; 28 +/- 4 mumol/L, controls). The activity appears to be a specific acetylhydrolase rather than a phospholipase A2 as preincubation of the substrate with 0 to 100 mumol/L phosphatidylcholine did not affect the amount of [3H] acetate released. The role of PAF, and its degradation by LDL-bound PAF acetylhydrolase in the phenotypic expression of this patient with Tangier disease, is not known. However, this is the first patient so far described who has an increased ability to degrade PAF in the plasma.
Demonstration of burst-promoting activity of recombinant human GM-CSF on circulating erythroid progenitors using an assay involving the delayed addition of erythropoietin
Authors: Donahue RE , Emerson SG , Wang EA , Wong GG , Clark SC , Nathan DG
Published: October 2019

We demonstrate through the use of an in vitro assay involving the delayed addition of erythropoietin that human recombinant GM-CSF, cloned from a mature T cell line, Mo, clearly has burst-promoting activity (BPA) on peripheral blood erythroid progenitors at picomolar concentrations. Delay for up to 72 hours of the addition of erythropoietin to semi-solid methylcellulose cultures of concentrated peripheral blood progenitors minimizes or eliminates BPA-independent erythroid colony formation with little loss of BPA-dependent erythroid colony formation. This assay will prove useful in accurately detecting sources of BPA.
Abnormal spectrin in hereditary elliptocytosis
Authors: Marchesi SL , Knowles WJ , Morrow JS , Bologna M , Marchesi VT
Published: October 2019

An abnormal alpha subunit of erythrocyte spectrin has been described in hereditary pyropoikilocytosis (HPP), a rare hemolytic anemia characterized by erythrocyte budding and fragmentation. In HPP spectrin, the N terminal domain of the alpha subunit (alpha I T80) shows increased susceptibility to tryptic digestion, resulting in cleavage to a 50,000-d peptide, presumably due to a change in primary structure of the alpha I domain which alters conformation and generates the new cleavage site. The functional result of this conformational alteration is marked impairment of spectrin oligomer formation in vitro, consistent with the established role of alpha I T80 in spectrin self-association. In the present study, we demonstrate an abnormal spectrin alpha subunit in two kindreds with hereditary elliptocytosis (HE) that is qualitatively identical to HPP spectrin. Clinical expression of HE in these families ranges from mild elliptocytosis without hemolysis to severe poikilocytic hemolytic anemia clinically resembling HPP. In all affected individuals, a fraction of alpha I T80 is abnormal, as shown by its cleavage during mild tryptic digestion to the 50 kd peptide described in HPP; the fraction of alpha I T80 affected is directly proportional to the severity of clinical expression of HE. Spectrin oligomer formation is likewise impaired to a degree which correlates with hematologic disease. One of the HE kindreds studied demonstrated polymorphism in the spectrin alpha II domain, previously described as a frequent occurrence in blacks. This family also demonstrates a variant alpha III domain in spectrin that has not previously been described. We conclude that the abnormality in the alpha I domain originally described in HPP spectrin is shared by a subset of patients with HE; the severity of clinical expression, ranging from mild nonhemolytic HE to poikilocytic hemolytic anemia, is related to the fractional quantity of the alpha subunit that is affected.
Chronic B-lymphocytic leukemia cells proliferate and differentiate following exposure to interferon in vitro
Authors: Ostlund L , Einhorn S , Robert KH , Juliusson G , Biberfeld P
Published: October 2019

The ability of interferon (IFN) to induce proliferation and differentiation in malignant B cells from 29 patients with chronic lymphocytic leukemia (CLL) and in lymphoid cells from 11 healthy donors was investigated. IFN induced transformation and plasmacytoid differentiation in B cells from 19 of 29 CLL patients. The transformed cells belonged to the malignant clone as indicated by the immunoglobulin (lg) light chain restriction. Cells exposed to IFN expressed intracellular lg to a varying degree, which was correlated to the level of plasmacytoid differentiation. IFN gave rise to proliferative responses in cells from three patients. Cytogenetic studies on lymphoid cells from one patient showed that proliferation occurred in the malignant B cells. Induction of proliferation and differentiation was observed with various alpha-IFN and gamma-IFN preparations, as well as with a completely pure beta-IFN, showing that IFN and not contaminants in the preparations were responsible for the observed effects. Maximal transformation and proliferation usually occurred after four days of incubation at an IFN concentration of 500 to 5,000 U/mL. The ability of IFN to induce differentiation in CLL cells may be of importance for the reported antitumoral effects observed in some B cell malignancies during IFN therapy.
Donor-derived red blood cell antibodies and immune hemolysis after allogeneic bone marrow transplantation
Authors: Hows J , Beddow K , Gordon-Smith E , Branch DR , Spruce W , Sniecinski I , Krance RA , Petz LD
Published: October 2019

Six cases of immune hemolytic anemia attributed to donor-derived red cell antibodies after allogeneic bone marrow transplantation (BMT) are reported. In 2/6 cases, severe intravascular hemolysis was seen, 6/6 required increased red cell transfusion, and 1/6 was treated by plasma exchange. All recipients were receiving cyclosporine to prevent graft-v- host disease. Investigations showed that in each case, the donor lacked ABO or Rho(D) red cell antigens present in the recipient. The direct antiglobulin test was positive in 6/6. Relevant serum antibody (anti-A, four cases; anti-B, one case; anti-D, one case) was first detected one to three weeks after BMT. Eluates made from recipient red cells showed the same specificity as serum antibody. Maximum hemolysis occurred nine to 16 days after BMT, suggesting that active production of antibody by “passenger” donor lymphocytes was the likely mechanism of hemolysis, rather than passive transfer of antibody in the marrow infusion. Retrospective analysis of 21 consecutive cyclosporine-treated BMT patients receiving marrow lacking ABO or D antigens present in the recipient showed that (1) 15/18 patients tested had red cell antibody production against recipient red cell antigens; (2) despite the frequent presence of antibody specific for recipient red cell antigens, only 3/21 patients developed clinically significant hemolysis; (3) clinical hemolysis could not be predicted by donor or recipient red cell antibody titers. We conclude that although red cell antibody against recipient antigens is frequently produced after minor ABO and D mismatched BMT in cyclosporine-treated recipients, only 10% to 15% of cases develop clinically significant immune hemolysis. The data presented show that the most likely source of antibody is “passenger” donor lymphoid cells.
The risks of central nervous system relapse and leukoencephalopathy in patients receiving marrow transplants for acute leukemia
Authors: Thompson CB , Sanders JE , Flournoy N , Buckner CD , Thomas ED
Published: October 2019

The records of 415 patients who received allogeneic marrow transplants for acute leukemia were reviewed to assess the risk of central nervous system (CNS) relapse and leukoencephalopathy after marrow transplantation. The Kaplan-Meier estimates of the probability of CNS relapse posttransplant were 13% for patients with acute lymphoblastic leukemia (ALL) and 2% for patients with acute nonlymphoblastic leukemia (ANL). Previous CNS disease was significantly correlated with an increased risk of CNS relapse in patients transplanted for ALL but not for ANL. In contrast, bone marrow involvement with leukemia at the time of transplant was associated with an increased risk of CNS relapse in patients with ANL but not in patients with ALL. Seventy-one patients with ALL did not receive posttransplant intrathecal methotrexate (IT- MTX) and 127 did. The probability of CNS relapse in these two groups was 38% and 7%, respectively (P less than .02). This protective benefit from IT-MTX was present in patients both with and without a history of CNS involvement or marrow involvement at the time of transplant. In patients with ANL, 116 patients did not receive posttransplant IT-MTX and 101 patients did, but no protection from CNS relapse was observed from IT-MTX irrespective of a patient's previous CNS history or marrow status at the time of transplant. Leukoencephalopathy was seen exclusively in patients who had received radiation and/or intrathecal chemotherapy to the CNS before preparation for marrow transplantation and posttransplant IT-MTX. In such patients the risk of leukoencephalopathy was 7%. From our data, it appears that posttransplant IT-MTX is a significant benefit for ALL patients in preventing CNS relapse after marrow transplantation. A similar benefit from posttransplant IT-MTX for ANL patients cannot be established from this study. In both groups, increasing total CNS therapy was associated with an increasing risk of leukoencephalopathy.
Lymphokine induction of NK-like cytotoxicity in T cells from B-CLL
Authors: Alvarez de Mon M , Casas J , Laguna R , Toribio ML , de Landazuri MO , Durantez A
Published: October 2019

T cells from patients with B cell chronic lymphocytic leukemia (B-CLL) exhibit defective natural killer (NK) activity. In this study, we have analyzed the cytotoxic-inducer effects of gamma interferon (gamma-IFN) and supernatants containing interleukin 2 (IL 2 sup). T cells from patients with B-CLL were incubated with gamma-IFN or IL 2 sup. gamma- IFN did not modulate the very low or undetectable levels of NK activity present in the T cell population. However, the IL 2 sup induced a potent cellular cytotoxicity against NK-sensitive and NK-resistant tumoral target cells. This cytotoxic inducer effect (a) was present in lectin-free IL 2 sup and in a 15,000- to 20,000-dalton molecular weight fraction obtained by gel filtration chromatography of this supernatant; (b) was directed against NK-sensitive and NK-resistant target cells; (c) was not correlated with the basal levels of NK activity; and (d) was not associated with a development or augmentation of the proportion of lymphocytes with classic NK cell phenotype. Taken together, these results demonstrate that unstimulated T cells from B-CLL patients, incubated briefly (18 hours) with IL 2 sup but not gamma-IFN, have strong NK-like cytotoxicity, despite the lack of classic NK activity.
Purification of human erythropoietin to homogeneity by a rapid five- step procedure
Authors: Krystal G , Pankratz HR , Farber NM , Smart JE
Published: October 2019

Human urinary erythropoietin (Ep) has been purified using a simple five- step procedure to yield preparations with potencies of 80,000 U/mg in 25% yield. The five steps involve: (1) affinity chromatography on CM Affi-Gel Blue, (2) chromatofocusing, (3) wheat germ lectin (or hydroxylapatite) chromatography, (4) reverse-phase high-performance liquid chromatography (HPLC) using a phenyl column, and (5) preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Ep activity was determined at each stage using a highly sensitive and specific in vitro assay that measures [3H]-thymidine incorporation into erythroid cells from spleens of phenylhydrazine-treated mice. The step 5 material was also tested with the in vivo polycythemic mouse assay procedure and was found to have a similar potency to that obtained in the [3H]-thymidine in vitro assay. SDS-PAGE analysis of the step 5 material revealed a single 38.5-kd band that comigrated with Ep bioactivity. Homogeneity was confirmed by amino acid sequence analysis. Starting with urine containing approximately 13 U/mg of protein, the cumulative degrees of purification achieved with each step were: step 1,25-fold; step 2, 75-fold; step 3, 300-fold; step 4, 1,500-fold; and step 5, 5,000-fold. Corresponding overall recoveries after each step were: greater than 100%, 70%, 45%, 30%, and 25%. These recoveries could be obtained when as little as 5,000 U of starting urinary Ep were processed because of the introduction of Tween 20 and SDS into buffers used at various stages of the purification procedure. In addition, a rapid method for determining Ep purity which involves reverse-phase HPLC of trypsinized 125I-labeled Ep is presented. This allows the establishment of purity with far less material than is required for amino acid sequencing.
Detection of clonal excess in lymphoproliferative disease by kappa/lambda analysis: correlation with immunoglobulin gene DNA rearrangement
Authors: Berliner N , Ault KA , Martin P , Weinberg DS
Published: October 2019

Previous studies have suggested that analysis of the distribution of surface immunoglobulin light chain isotypes by flow cytometry provides evidence for monoclonality of B cell tumors and may detect populations of circulating tumor cells in patients with lymphoproliferative disease. We have used simultaneous flow cytometry and DNA restriction enzyme analysis on 58 samples of tissue and blood to determine whether lymphocyte populations detected by “kappa/lambda” analysis are indeed monoclonal. In greater than 90% of cases, abnormalities detected by flow cytometry correlated with monoclonal rearrangements of immunoglobulin genes as detected by Southern blot analysis. By analyzing tissue and blood from the same patients, we have also demonstrated that monoclonal circulating cells detected by flow cytometry reflect peripheral circulating tumor cells, since DNA from these cells shows the same immunoglobulin rearrangement as DNA from the original tumors in these patients. Although mixing studies suggested that DNA rearrangement studies were more sensitive than was flow cytometry in detecting minor populations of monoclonal lymphocytes, we found only one case in which this affected the diagnostic accuracy of the kappa/lambda analysis, with one notable exception, that of detection of a monoclonal proliferation of B cells that did not express surface immunoglobulin. The kappa/lambda test thus offers a powerful diagnostic tool in the evaluation of lymphoproliferative disease.
Treatment of multiple myeloma with recombinant alpha-interferon
Authors: Quesada JR , Alexanian R , Hawkins M , Barlogie B , Borden E , Itri L , Gutterman JU
Published: October 2019

Thirty-two patients with multiple myeloma were treated with recombinant alpha-interferon clone A (rIFN alpha A) daily by intramuscular injection with an initial dose of 12 X 10(6) U/m2. Of 27 patients evaluable for response, tumor responses were obtained in seven of 14 previously untreated patients (50%) and two of 13 who had relapsed or failed prior chemotherapy (15%). In all patients who had tumor response, there was restoration from subnormal levels of serum immunoglobulins, an effect infrequently observed with chemotherapy. The median duration of tumor responses exceeded 14 months (range, 6 to 20). Moderate-to-severe fatigue was the predominant side effect and necessitated dose reductions in all patients. We conclude that treatment of early stages of multiple myeloma with rIFN alpha A is beneficial because of the substantial response rate and the improvement in the synthesis of serum immunoglobulins. rIFN alpha A has a potential role in combination with other agents in the treatment of multiple myeloma.
Immunoregulation of B lymphocyte colony formation by T cell subsets in patients with chronic lymphocytic leukemia
Authors: Fernandez LA , MacSween JM , Langley GR
Published: October 2019

Normal B lymphocytes are activated, proliferate, and then differentiate into plasma cells and secrete immunoglobulin (Ig). We have reported that chronic lymphocytic leukemia (CLL) T4 cells help and CLL T8 cells lack suppressor effects on Ig synthesis by normal B cells (Blood 62:767, 1983). We have now explored the earlier phase, proliferation, using B cell colony formation; in semisolid media. B lymphocyte colonies from normal individuals and from patients with CLL were grown in 0.3% agarose overlayed with T cells or T cell subsets and the B cell mitogen staphylococcal protein A. Enriched T cells, OKT4 or OKT8, were obtained either by sheep erythrocyte rosettes or depletion of OKT8 or OKT4 cells by monoclonal antibody or complement, respectively. Twenty thousand B cells from normal subjects yielded 65 +/- 9, 64 +/- 7, and 19 +/- 6 colonies with autologous unfractionated T-, OKT4-, or OKT8- positive cells, respectively. This compared to 29 +/- 11, 81 +/- 11, and 15 +/- 4 colonies from patients with CLL with added autologous unfractionated T-, OKT4-, or OKT8-positive cells. To determine whether the fewer number of colonies in both normal subjects and patients with CLL with OKT8-positive cells was due to suppression or lack of help, the number of OKT4-positive cells was held constant, and OKT8-positive cells were added in increasing numbers. No suppression of colony formation could be demonstrated. Furthermore, the addition of increasing numbers of concanavalin A (Con A)-activated OKT8-positive cells did not suppress colony formation. These results suggest that the CLL T cell subsets behave in a functionally similar manner to normal T cell subsets, namely, (1) that normal and CLL B cell colony growth is helped by OKT4 cells; and (2) in contrast to immunoglobulin secretion by B cells, neither normal nor CLL OKT8 cells, unstimulated or activated by Con A, suppress B cell colony growth.
An ultrastructural morphometric analysis of platelet giant and fusion granules
Authors: Payne CM , Glasser L
Published: October 2019

Our purpose in this study was to establish ultrastructural morphometric criteria that can be used to define pathologic giant and fusion platelet granules and to determine whether patients with neoplastic myeloproliferative disorders (MPD) can be distinguished from other patients. We have morphometrically analyzed 2,391 giant and fusion granule profiles from 46 patients with neoplastic MPD, 127 other diseased control subjects, and 30 normal subjects using a computerized image analyzer. The largest granule profile observed in normal subjects had an area of 0.51 mu 2 and a perimeter of 3.21 mu. The most irregularly shaped of the large granule profiles photographed from normal subjects had a form factor (FF) value of 0.31. FF values indicate the degree of deviation of a given granule contour from a circle and is expressed by the formula 4 pi A/P2. A pathologic granule profile was then defined as a granule that exceeded any of these limits. Fifty-seven percent of the patients with neoplastic MPD were determined to have abnormal platelet granule profiles using the aforementioned morphometric criteria. It was determined, however, that morphometrically defined giant- and fusion-type granule profiles were nonspecific and were also found in 20% of the other diseased patients with no clinical evidence of an underlying neoplastic MPD. Morphometry has allowed us to define the upper limits of normal for the area and perimeter of individual platelet granules. Morphometrically defined giant fusion granules were determined to be more prevalent in the neoplastic MPD group, but because of their nonspecificity, may only have diagnostic significance for the individual patient in specific clinical settings. The pathogenesis of platelet fusion granules is discussed.
Somatic shift to homozygosity for a chromosomal polymorphism in a child with acute lymphoblastic leukemia
Authors: Stamberg J , Shende A , Lanzkowsky P
Published: October 2019

A 12-year-old girl with acute lymphoblastic leukemia (ALL) had two types of acquired cytogenetic abnormalities in her pretreatment peripheral blood and bone marrow: hyperdiploidy due to tetrasomy 8, 10, and 21; and, in the hyperdiploid cells, a shift from heterozygosity to homozygosity for a polymorphic variant on chromosome 15. Both abnormalities disappeared after chemotherapy, when the patient entered clinical remission. It has recently been found that shifts to homozygosity occur in retinoblastoma and Wilms' tumor. Our observation extends this finding to leukemia and indicates that such shifts may have general importance in tumorigenesis.
Synergistic effects of butyrate on platelet responses to arachidonate, A23187, PGE1, and forskolin
Authors: Tandon NN , Tralka TS , Jamieson GA
Published: October 2019

With eukaryotic cells, butyrate is known to induce a series of morphological and biochemical changes that mimic cellular differentiation. With platelets, we have found that butyrate (10 mmol/L) caused an approximately threefold increase in sensitivity to calcium ionophore A23187 and arachidonate. Maximum aggregation was observed at agonist concentrations of 3 mumol/L and 170 mumol/L, respectively, as compared with required concentrations of 10 mumol/L and 400 mumol/L in the absence of butyrate. Similar effects were seen with isobutyric acid, and about one-half the effect was shown with valerate and caproate, but lower homologues showed no synergistic effect. No ultrastructural changes were observed in platelets incubated with butyrate, and the aggregation effects were reversible and returned to normal on removal of butyrate. Membrane fluidity was unchanged by butyrate as measured by changes in the fluorescence depolarization of diphenylhexatriene. Butyrate caused a 60% to 70% increase in the uptake of 3H-arachidonate. Butyrate also potentiated the inhibition of platelet function by prostaglandin E1 and forskolin and uptake of 3H- forskolin was increased approximately 20%. In contrast, platelet response to other agonists (ADP, epinephrine, collagen, thrombin, and platelet-activating factor) was essentially unaffected by butyrate. These results suggest that butyrate may increase the uptake of certain hydophobic agonists and antagonists by platelets. Similar mechanisms for uptake of endogenous effectors may explain the response of eukaryotic cells to butyrate in culture.
Altered stem cell (CFU-S) function following infection of hematopoietic cells with a virus carrying V-src
Authors: Boettiger D , Dexter TM
Published: October 2019

Long-term murine bone marrow cultures were used to support the growth and development of hematopoietic cells. After hematopoiesis was established, the cultures were infected with a recombinant murine amphotropic virus carrying the avian sarcoma virus src gene and the CFU- S kinetics were examined. The CFU-S from the src-infected cultures displayed a reduced seeding efficiency in the standard spleen colony assay. The self-renewal capacity of these CFU-S was tested by their ability to reestablish hematopoiesis when serially transplanted on irradiated bone marrow cultures and by serial passage in spleens of irradiated mice. In both tests, cells from the src-infected cultures exhibited an enhanced ability to sustain a high level of self-renewal. The other property of stem cells which may be measured is the probability of self-renewal at each cell division which dictates the distribution between stem cells and differentiated type progeny. CFU-S from the src-infected cultures had higher average probabilities of self- renewal and therefore reduced differentiation. These differences suggest that expression of src had indirectly or directly altered the normal differentiation program of the stem cells.
Effect of leukocyte antibodies and HLA matching on the intravascular recovery, survival, and tissue localization of 111-indium granulocytes
Authors: McCullough J , Clay M , Hurd D , Richards K , Ludvigsen C , Forstrom L
Published: October 2019

The effect of leukocyte antibodies detected under different conditions on the fate in vivo of granulocytes was studied using 111-indium- labeled granulocytes. Sera from patients were tested by granulocyte agglutination (GA), granulocytotoxicity (GC), granulocyte immunofluorescence (GIF), lymphocytotoxicity (LC), and antibody- dependent lymphocyte-mediated granulocytotoxicity. Granulocytes from donors to be studied were labeled with 111-indium and injected. Then the intravascular recovery and survival or tissue localization was determined in 93 studies. Antibodies detected by granulocyte agglutination were associated with a significant reduction in recovery (6.7% v 30.8% in controls; P less than .001) and t1/2 (0.3 hours v 5.6 hours in controls; P = .002). When all possible combinations of serum reactivity were considered, reactivity in the GA plus GIF assays had the best correlation with decreased recovery (R2 = .49; P less than .001) and t1/2 (R2 = .73; P less than .001). When the relationship between the strength of antibody reactivity and the recovery and t1/2 were analyzed, the best relationship was between the combination of LC and GIF with recovery (R2 = .62; P = .001). Because of the general availability of the HLA (LC) testing, the role of LC reactivity was investigated in other ways. There was a strong relationship between sera highly reactive by LC and those reactive by GIF. These highly reactive sera were also associated with reduced recovery and t1/2. The influence of specific HLA antigen mismatches was also studied. When donor and recipient were mismatched for the HLA-A2, B8, or BW44 antigens, there was a significant reduction in either recovery, t1/2, or both. Tissue localization was studied by body scans in patients with and without known sites of inflammation. Antibodies detected by a combination of GA and GIF caused abnormal pulmonary sequestration of granulocytes (three cases) and failure of granulocytes to localize at known sites of inflammation (three cases). HLA (LC) antibodies did not alter tissue localization despite the presence of the corresponding HLA antigens on granulocytes. It appears that GA, GIF, or a combination of these tests is the most effective predictor of altered in vivo fate of granulocytes. However, sera highly reactive by LC and GIF probably define a group of highly immunized patients in whom granulocyte recovery and t1/2 are also reduced. Mismatching for certain HLA antigens is also associated with reduced granulocyte recovery and survival. At present, GA, with or without the immunofluorescence assay, is the most effective predictor of altered in vivo granulocyte activity.(ABSTRACT TRUNCATED AT 400 WORDS)
A novel mutation in the TATA box in a Japanese patient with beta +- thalassemia
Authors: Takihara Y , Nakamura T , Yamada H , Takagi Y , Fukumaki Y
Published: October 2019

A single base substitution (A-G) at position -31 within the highly conserved proximal promoter element, the TATA box, was identified in the beta-globin gene cloned from a Japanese woman with beta +- thalassemia. It appears that she is homozygous for this specific allele, as determined by haplotype analysis using seven different polymorphic sites in the beta-globin gene cluster. Transient expression of the mutant gene in COS cells revealed a 45% reduction in beta-globin RNA production, relative to normal. These results establish the functional significance of the second base of the TATA box for in vivo transcription of the human beta-globin gene.
Thrombospondin in essential thrombocythemia
Authors: Lawler J , Cohen AM , Chao FC , Moriarty DJ
Published: October 2019

Essential thrombocythemia is a myeloproliferative disorder characterized by frequent bleeding and thrombotic complications. On a molecular level, two abnormalities of platelet thrombospondin have been identified: abnormal glycosylation of the intact 185,000-dalton chain has been detected and a shortened form of the thrombospondin chain is present. We have used two monoclonal antibodies and Lens culinaris lectin to probe the structure of thrombospondin in the platelets from three patients with essential thrombocythemia; one patient with polycythemia vera and two patients with secondary thrombocytosis. The presence of abnormal thrombospondin fragments with molecular weights of 160,000 and 30,000 was detected in the intact platelets and in the supernatant from thrombin-treated platelets, in all of the individuals except one of the secondary thrombocytosis patients. Monoclonal antibody binding studies indicate that both fragments are produced by proteolysis at a single site, which results in the removal of a 30,000- dalton fragment from the NH2-terminal. Lens culinaris lectin-binding studies revealed that some of the carbohydrate moieties of thrombospondin are near this cleavage site. The results are consistent with the hypothesis that the abnormal thrombospondin fragments observed under conditions of increased platelet production are due to increased susceptibility to proteolysis which, in turn, may be due to defective glycosylation.
Production of B cell growth factor(s) by neoplastic B cells from hairy cell leukemia patients
Authors: Ford RJ , Kwok D , Quesada J , Sahasrabuddhe CG
Published: October 2019

Recent studies have shown that normal human T cells contain a high- molecular-weight (mol wt) protein exhibiting B cell growth factor (BCGF) activity. Other studies have shown that virally transformed human B cells also secrete a high-mol-wt BCGF-like molecule in vitro. We have studied neoplastic B cells from patients with untreated hairy cell leukemia (HCL) to ascertain whether such cytoplasmic BCGF activity is present in the tumor cells. Studies on HCL cells from four patients indicated that BCGF-like activity was in fact present in the cytosolic extracts when tested on autochthonous HCL cells as well as on normal BCGF-dependent human B cell lines. Chromatographic analysis indicated that the BCGF activity from HCL cells was similar in mol wt as well as function to the normal T cell-derived cytosolic BCGF activity. These studies suggest that HCL cells contain and, in some cases, secrete a high-mol-wt growth factor that can be autostimulatory and appears to resemble a similar growth factor molecule found in normal human T cells.
Defective T cell responsiveness in chronic lymphocytic leukemia: analysis of activation events
Authors: Kay NE , Kaplan ME
Published: October 2019

Chronic lymphocytic leukemia (CLL) is a B cell disorder in which major T cell proliferative defects are present. We investigated the nature of this deficit by studying several parameters known to be crucial in normal T cell proliferative response to mitogen. Purified peripheral blood T cells from B-CLL patients were analyzed for the presence of T3 antigen. We observed that CLL T cells have a direct correlation between levels of T3 membrane antigen and proliferative response to mitogen. The appearance of activation antigens (transferrin, HLA-DR, and interleukin 2 [IL 2] receptor) was normal in CLL T cells post-mitogen exposure. Despite the normal presentation of IL 2 receptor on CLL T cell membrane, there was decreased production of IL 2 by CLL patients (v controls) (39.6 +/- 10.2 cells per milliliter v 64.6 +/- 11.0 cells per milliliter). Finally, we were able partially, but not fully, to reconstitute CLL T proliferative response to mitogen by adding purified exogenous IL 2. These findings suggest that CLL T cells have multiple defects that may impact on their proliferative potential. Further insight into these deficits may result in strategies that will facilitate immunologic restoration in T cells of these patients.
Iron metabolism in the Belgrade rat
Authors: Edwards J , Huebers H , Kunzler C , Finch C
Published: October 2019

Iron metabolism in the Belgrade rat was examined in the intact animal and in the reticulocyte suspensions. The plasma iron turnover was increased. However, when allowance was made for the effect of the elevated plasma iron concentration, erythroid marrow capacity for iron uptake was at basal levels. Numbers of erythroid cells in marrow and spleen measured by the radioiron dilution technique were increased. Thus iron uptake was not proportionate to the erythroid hyperplasia in the b/b rat, despite a more than adequate plasma iron supply. This relative deficiency in iron uptake was reflected in a severe microcytosis and elevated red cell protoporphyrin. Reticulocyte incubation studies demonstrated an unimpaired uptake of the transferrin- iron-receptor complex but a marked reduction in iron accumulation. The diferric transferrin molecule, when it did give up iron within the cell, released both of its iron atoms so that only apotransferrin was returned to the media. In contrast to the nearly complete release of iron within the normal reticulocyte, the major portion of iron taken up by the Belgrade reticulocyte was returned to the plasma. The release mechanism that can be impaired in iron-deficient reticulocytes by EDTA or cadmium was shown to be affected by lower concentrations of these substances in the Belgrade reticulocyte. It is concluded that the Belgrade rat has an abnormality of iron release within the absorptive vacuole that is responsible for a state of intracellular iron deficiency, involving the erythron and other body tissues.
The sensitivity to cytosine arabinoside of the blast progenitors of acute myeloblastic leukemia
Authors: Nara N , Curtis JE , Senn JS , Tritchler DL , McCulloch EA
Published: October 2019

Two culture methods are available for the study of the blast cells of acute myeloblastic leukemia (AML). One is an assay for clonogenic precursors; it depends on their ability to form blast colonies in culture in the presence of methylcellulose and suitable growth factors. The other assesses the growth of blast cells in suspension culture, where growth is measured by increasing numbers of clonogenic cells. We have compared the two methods as assays for the cytotoxic effects of the chemotherapeutic drug cytosine arabinoside (Ara-C). Marked patient- to-patient variation was found using either method; however, the slopes of the dose-response curves were usually greater when cells were exposed to drug in suspension rather than in methylcellulose. Control experiments showed that the difference could not be explained by drug carry-over from the suspension cultures to the methylcellulose plates when clonogenic cells in the suspensions were assessed. Further, the survival curves for Adriamycin were very similar, regardless of which assay was used. No correlation was found between D10 Ara-C values measured in suspension or in methylcellulose. However, a significant association with outcome was found between D10 Ara-C in suspension and response to treatment with a regimen in which Ara-C was the only chemotherapeutic agent used. No such association was detected when the D10 values obtained with the clonogenic assay were compared with outcome for the same group of 15 patients. Finally, a feasibility experiment was performed in which blast cells were exposed to Ara-C repeatedly during exponential growth over 238 days. A dose-related inhibition of growth was observed; no evidence was seen of emerging drug-resistant cells. Nor did the morphology of the cells change as a result of drug exposure. We conclude that drug sensitivities of AML blast cells in culture are dependent on measurement methods, even when techniques affecting cell proliferation are compared. Measurements of drug sensitivity in culture may best be interpreted when the bases of the assay systems are understood.
Diversity of human gamma-globin gene loci including a quadruplicated arrangement
Authors: Shimasaki S , Iuchi I
Published: October 2019

A quadruplicated gamma-globin gene as a (5′-G gamma-gamma-gamma-A gamma- 3′) was detected in an adult Chinese during a survey designed to detect G gamma- and A gamma-globin genes in Japanese. Five triplicated (-G gamma-gamma-A gamma-) and two single (-gamma-) haplotypes were also detected in 103 healthy adult Japanese. All of the unusual chromosomes appeared to reflect an unequal but homologous crossover between G gamma- and A gamma-globin genes. A new Bgl II polymorphic site located around the 3′ terminal region of the G gamma-globin gene was also discovered.
Abnormal in vitro differentiation of peripheral blood clonogenic B cells in common acute lymphoblastic leukemia during complete remission
Authors: Consolini R , Breard J , Bourinbaiar A , Goutner A , Georgoulias V , Canon C , Brugerie E , Mathe G
Published: October 2019

An in vitro B cell colony assay system was used to evaluate B cell growth from peripheral blood precursors in common acute lymphoblastic leukemia (CALL) patients in remission during maintenance therapy and in normal controls. Major differences between the two groups were found in the phenotypic and morphologic features of pooled colony cells. In both cases, the cells were E-. Controls' cells were surface immunoglobulin (sIg)-positive, and some (mean, 25%) expressed la determinants. By Wright-Giemsa staining, they appeared as plasmacytoid cells. In contrast, patients' cells had predominantly a lymphoblastoid appearance, fewer cells had developed sIg, and a large fraction (mean, 43%) were Ia-positive. Moreover, the CALL antigen (CALLA) was expressed by a mean of 18% (range, 2% to 72%) of the patients' colony cells, whereas CALLA was never found in control colonies. Thus, cells with immature features persist in the colonies of CALL patients. Secondary colonies could be generated from the patients' cultured cells, indicating their self-renewal capacity. CALLA + cells were also present in the secondary colonies. Finally, cytogenetic studies showed that a fraction of the patients' colony cells had karyotypic abnormalities similar to that of the original lymphoblasts. It is believed that in CALL patients this B cell assay permits the clonal expansion of residual circulating cells linked to malignant clones that are not detectable by classic hematologic and cytologic methods.