bluuebunny/crossref_metadata_2025_split huggingface dataset:
OX40 Expressed on Fresh Leukemic Cells From Adult T-Cell Leukemia Patients Mediates Cell Adhesion to Vascular Endothelial Cells: Implication for the Possible Involvement of OX40 in Leukemic Cell Infiltration
Authors: Imura Akihiro , Hori Toshiyuki , Imada Kazunori , Kawamata Shin , Tanaka Yuetsu , Imamura Sadao , Uchiyama Takashi 
Published: October 2019

AbstractWe demonstrated previously that OX40 and its ligand, gp34, directly mediate adhesion of activated normal CD4+ T cells, as well as human T-cell leukemia virus type I (HTLV-I)–transformed T cells to vascular endothelial cells. In the present study, we examined expression of OX40 on fresh leukemic cells from patients with adult T-cell leukemia (ATL) and its possible involvement in cell adhesion. Flow cytometric analysis showed that peripheral blood mononuclear cells (PBMC) or lymph node tumor cells from 15 of 17 cases expressed significant levels of OX40 without stimulation. On the other hand, gp34 was not expressed on these cells, although its expression is also known to be associated with HTLV-I-infection. In Western blot analysis, a 50-kD protein band was detected by anti-OX40 monoclonal antibody (MoAb) in two ATL cases examined, as well as phytohemagglutinin (PHA) blasts and Hut102, an HTLV-I–infected T-cell line, but not in resting PBMC or Jurkat. Expression of OX40 mRNA was shown by reverse transcriptase-polymerase chain reaction in all ATL cases tested, PHA-blasts, and Hut102, but not in resting PBMC or Jurkat. We could not detect expression of HTLV-I viral mRNA in any of the cases tested. Cell adhesion assay was performed and in at least three cases, fresh ATL cells exhibited adhesion to human umbilical vein endothelial cells that could be considerably inhibited by either anti-OX40 MoAb or anti-gp34 MoAb. Immunohistochemical staining of skin biopsy specimens indicated that infiltrating mononuclear cells express OX40 in vivo. Taken together, these data indicate that leukemic cells from most, but not all, ATL patients constitutively express OX40, which may play a role in leukemic cell infiltration in addition to cell adhesion in vivo.
Activation of the Stress-Activated Protein Kinases by Multiple Hematopoietic Growth Factors With the Exception of Interleukin-4
Authors: Foltz Ian N. , Schrader John W. 
Published: October 2019

AbstractThe stress-activated protein/c-Jun N-terminal kinases (SAPK/JNK) have been shown to be activated by pro-inflammatory cytokines, as well as physical and chemical stresses. We now show that a variety of hematopoietic growth factors, including Steel locus factor (SLF ), granulocyte-macrophage colony-stimulating factor (GM-CSF ), and interleukin-3 (IL-3), all of which promote the growth and survival of various lineages of hematopoietic cells, activate the stress-activated protein kinases in the factor-dependent cell line MC/9. These hematopoietic growth factors activated both 46- and 55-kD isoforms of both SAPKγ and SAPKα. Furthermore, we demonstrate that SAPK activation correlated with the phosphorylation of SAPK/ERK kinase-1 (SEK1) after treatment with SLF or GM-CSF. Interestingly, IL-4, a cytokine with distinctive and important effects on the immune system, was the exception among the hematopoietic growth factors we examined in failing to induce activation of SAPKγ, SAPKα, or SEK1. These findings show that activation of SAPK is involved, not only in responses to stresses, but also in signaling by growth factors that regulate the normal development and function of cells of the immune system.
L-Selectin–Dependent Leukocyte Adhesion to Microvascular But Not to Macrovascular Endothelial Cells of the Human Coronary System
Authors: Zakrzewicz A. , Gräfe M. , Terbeek D. , Bongrazio M. , Auch-Schwelk W. , Walzog B. , Graf K. , Fleck E. , Ley K. , Gaehtgens P. 
Published: October 2019

AbstractTo characterize L-selectin–dependent cell adhesion to human vascular endothelium, human cardiac microvascular endothelial cells (HCMEC) and human coronary endothelial cells (HCEC) were isolated from explanted human hearts. The adhesion behavior of human (NALM-6) and mouse (300.19) pre-B cells transfected with cDNA encoding for human L-selectin was compared with that of the respective nontransfected cells in a flow chamber in vitro. More than 80% of the adhesion to tumor necrosis factor-α (TNF-α)–stimulated HCMEC at shear stresses <2 dyne/cm2 was L-selectin dependent and could be equally well blocked by an anti–L-selectin antibody or a L-selectin-IgG-chimera. No L-selectin dependent adhesion to HCEC could be shown. The L-selectin dependent adhesion to HCMEC was insensitive to neuraminidase, but greatly inhibited by addition of NaClO3 , which inhibits posttranslational sulfation and remained elevated for at least 24 hours of stimulation. E-selectin dependent adhesion of HL60 cells to HCMEC was blocked by neuraminidase, but not by NaClO3 and returned to control levels within 18 hours of HCMEC stimulation. It is concluded that microvascular, but not macrovascular endothelial cells express TNF-α–inducible sulfated ligand(s) for L-selectin, which differ from known L-selectin ligands, because sialylation is not required. The prolonged time course of L-selectin dependent adhesion suggests a role in sustained leukocyte recruitment into inflammatory sites in vivo.
Differential Effects of Interleukin-13 on Cytomegalovirus and Human Immunodeficiency Virus Infection in Human Alveolar Macrophages
Authors: Hatch William C. , Freedman Andrew R. , Boldt-Houle Deborah M. , Groopman Jerome E. , Terwilliger Ernest F. 
Published: October 2019

AbstractAlveolar macrophages, which form a principal line of defense against a variety of pulmonary pathogens, may themselves be infected by viruses like human immunodeficiency virus-1 (HIV-1), which impair their defensive functions. Interleukin-13 (IL-13), a multifunctional cytokine, has been considered for therapeutic use based on its potent inhibition of HIV-1 in these cells. We have further examined the effects of IL-13 on alveolar macrophages under conditions that reflect those seen in acquired immune deficiency syndrome, where this cell type is often infected by the opportunistic pathogen human cytomegalovirus (HCMV). Alveolar macrophages exposed to both HCMV and HIV-1 consistently exhibited higher levels of HIV-1 replication than cells exposed to HIV-1 alone. HIV-1 production was strongly suppressed in alveolar macrophages treated with IL-13 regardless of whether or not the cultures were coinfected with HCMV. However, IL-13 treatment markedly enhanced the expression of HCMV in otherwise latently infected macrophages in a dose dependent manner. These unexpected differential effects of IL-13 on host-virus interactions are important considerations in guiding its potential therapeutic applications.
Studies on Abnormal Hemoglobins
Authors: SINGER KARL , CHAPMAN A. ZERNE , GOLDBERG SEYMOUR R. , RUBINSTEIN HERBERT M. , ROSENBLUM SOL A. 
Published: October 2019

Abstract
Four Negro patients with pure (homozygous) hemoglobin C disease are described, who exhibited the characteristic features of, (1) a hemolytic process with or without anemia, (2) splenomegaly, and, (3) numerous target cells in the film. The method of choice to establish the diagnosis objectively is paper electrophoresis of the stroma-free hemolysate, prepared from the patient’s erythrocytes. No alkali resistant (F) hemoglobin was found in this syndrome (homozygous for hemoglobin C) in contradistinction to sickle cell anemia (homozygous for hemoglobin S) where F hemoglobin is usually present. Splenectomy did not ameliorate the hemolytic process.
Incorporation of Radioactive L-Cystine and L-Methionine by Leukemic Leukocytes in Vitro
Authors: WEISBERGER AUSTIN S. , SUHRLAND LEIF G. , GRIGGS ROBERT C. 
Published: October 2019

Abstract
A method is described for studying the incorporation of radioactive amino acids by leukemic leukocytes in vitro. The effects of time of incubation, concentration of amino acid, temperature, pH, oxygen tension, electrolytes, and glucose concentration on the uptake of S35 L-cystine and S35 L-methionine were studied. The in vitro incorporation of these amino acids appears to be an active process and not solely dependent upon simple diffusion. Reproducible results were obtained under constant conditions. The methods are suitable for studying possible therapeutic agents as well as biochemical abnormalities of leukemic leukocytes.
Development of a Novel Selective Amplifier Gene for Controllable Expansion of Transduced Hematopoietic Cells
Authors: Ito Keiko , Ueda Yasuji , Kokubun Masaki , Urabe Masashi , Inaba Toshiya , Mano Hiroyuki , Hamada Hirofumi , Kitamura Toshio , Mizoguchi Hideaki , Sakata Tsuneaki , Hasegawa Mamoru , Ozawa Keiya 
Published: October 2019

AbstractTo overcome the low efficiency of gene transfer into hematopoietic cells, we developed a novel system for selective expansion of transduced cells. To this end, we constructed a chimeric cDNA (GCRER) encoding the fusion protein between the granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone-binding domain (HBD) of the estrogen receptor (ER) as a selective amplifier gene. Use of the intracellular signaling pathway of G-CSFR was considered to be appropriate, because G-CSF has the ability not only to stimulate the neutrophil production, but also to expand the hematopoietic stem/progenitor cell pool in vivo. To activate the exogenous G-CSFR signal domain selectively, the estrogen/ER-HBD system was used as a molecular switch in this study. When the GCRER gene was expressed in the interleukin-3 (IL-3)–dependent murine cell line, Ba/F3, the cells showed IL-3–independent growth in response to G-CSF or estrogen. Moreover, the Ba/F3 cells transfected with the Δ(5-195)GCRER, whose product lacks the extracellular G-CSF–binding domain, did not respond to G-CSF, but retained the ability for estrogen-dependent growth. Further, murine bone marrow cells transduced with the GCRER or Δ(5-195)GCRER gene with retroviral vectors formed a significant number of colonies in response to estrogen, as well as G-CSF, whereas estrogen did not stimulate colony formation by untransduced murine bone marrow cells. It is noteworthy that erythroid colonies were apparently formed by the bone marrow cells transduced with the GCRER gene in the presence of estrogen without the addition of erythropoietin, suggesting that the signals from the G-CSFR portion of the chimeric molecules do not preferentially induce neutrophilic differentiation, but just promote the differentiation depending on the nature of the target cells. We speculate that when the selective amplifier genes are expressed in the primitive hematopoietic stem cells, the growth signal predominates and that the population of transduced stem cells expands upon estrogen treatment, even if some of the cells enter the differentiation pathway. The present study suggests that this strategy is applicable to the in vivo selective expansion of transduced hematopoietic stem cells.
p16INK4A Promotes Differentiation and Inhibits Apoptosis of JKB Acute Lymphoblastic Leukemia Cells
Authors: Urashima Mitsuyoshi , DeCaprio James A. , Chauhan Dharminder , Teoh Gerrard , Ogata Atsushi , Treon Steven P. , Hoshi Yasutaka , Anderson Kenneth C. 
Published: October 2019

Abstract
Homozygous p16INK4A (p16) gene deletion is frequent in primary tumor cells from acute lymphoblastic leukemia (ALL), suggesting that loss of p16 may be an important precursor to transformation in ALL. We have previously described JKB, a human ALL cell line, that contains homozygous deletion of the p16 gene. Because ectopic expression of p16 suppresses cell growth, we created a temperature sensitive p16 mutant to develop a system for inducible p16 function in human ALL. JKB cells were transfected either with a p16 gene mutated at position 119 (E119G) to confer temperature sensitivity (JKB p16MT) or with control vector. The percentage of cells in G1 phase was similar in JKB control cells or in JKB p16MT cells cultured at restrictive conditions (40°C). However, with lowering of temperature from 40°C to permissive conditions (31°C), the percentage of JKB p16MT cells in G1 phase and binding of p16 to CDK4 and CDK6 increased, with associated decreases in CDK4 and CDK6 kinase activities, and dephosphorylation of retinoblastoma protein (pRB). Culture of JKB p16MT cells at 31°C for ≥3 days irreversibly inhibited growth. Moreover, JKB p16MT cells cultured under these permissive conditions showed a less transformed morphology and more differentiated phenotype than did these cells cultured under restrictive temperatures. Finally, dexamethasone (Dex) induced apoptosis of JKB p16MT cells cultured at 40°C, but did not trigger death of these cells cultured at 31°C. These results suggest that deletion of p16 gene in JKB human ALL cells is associated with dysregulated growth of less differentiated tumor cells, which nonetheless remain susceptible to apoptosis triggered by Dex.
The Exon 46-Encoded Sequence Is Essential for Stability of Human Erythroid α-Spectrin and Heterodimer Formation
Authors: Wilmotte Rick , Harper Sandra L. , Ursitti Jeanine A. , Maréchal Joëlle , Delaunay Jean , Speicher David W. 
Published: October 2019

AbstractHuman erythroid α-spectrin alleles responsible for hereditary elliptocytosis (αHE alleles) undergo increased incorporation into red blood cell membranes when the polymorphism αLELY (LELY: Low Expression LYon) occurs in trans. The αLELY polymorphism is characterized by a mutation in exon 40 at codon 1857 (CTA → GTA, Leu → Val) and the partial (50%) skipping of exon 46, which encodes residues 2177-2182 (Wilmotte et al, J Clin Invest 91:2091, 1993). Both of these peptide sequence alterations are located within the region of the α-chain involved in initiating heterodimer assembly, and either or both mutations could potentially contribute to decreased incorporation of α-chains from the αLELY allele in heterozygotes into red blood cell membranes. These possibilities were evaluated by testing the protease resistance and in vitro binding properties of normal and mutant recombinant 4-motif α subunit peptides containing the dimer initiation region. The two forms of α spectrin produced by alternative mRNA splicing of the αLELY allele were represented by α18-211857, a peptide with the codon 1857 mutation and retaining the exon 46 encoded sequence, and α18-211857-Δ46, a peptide carrying both the 1857 codon mutation and the exon 46 deletion. The properties of these two recombinant peptides were compared with α18-21, a peptide with the normal sequence at codon 1857 and retaining the exon 46 encoded sequence. The codon 1857 mutation does not adversely affect dimer formation, but it is responsible for the increased trypsin cleavage between the αIV and αV domains that was the characteristic feature initially used to identify the αLELY (SpαV/41) polymorphism (Alloisio et al, J Clin Invest 87:2169, 1991). Deletion of the six amino acids encoded by exon 46 perturbs folding of the α21 motif, because this region of the α18-211857-Δ46 peptide is rapidly degraded and this recombinant peptide is unusually prone to self-aggregation. Exon 46 deletion reduces, but does not eliminate, dimerization. Comparison of mild trypsin proteolytic products from an αLELY homozygote and the two αLELY recombinant peptides strongly suggests that little, if any, of the 50% of the α chains from the αLELY allele that contain the exon 46 deletion are incorporated into the mature erythroid membrane. Based on the in vitro analysis of recombinant αLELY peptides, the inability of detectable amounts of exon 46− α chains to assemble into the mature membrane skeleton in vivo is probably due to a combination of decreased dimer binding affinity and increased proteolytic degradation of these mutant chains.
Mutations in the Murine Fitness 1 Gene Result in Defective Hematopoiesis
Authors: Potter Mark D. , Shinpock Sarah G. , Popp Raymond A. , Popp Diana M. , Godfrey Virginia , Carpenter Donald A. , Bernstein Alan , Johnson Dabney K. , Rinchik Eugene M. 
Published: October 2019

AbstractIdentification and characterization of mutations that disrupt normal hematopoiesis are essential for understanding the genetic pathways that control the development and regulation of the mammalian hematopoietic system. Previously, the fitness 1 gene was identified by five, independent mutations in N-ethyl-N-nitrosourea (ENU) saturation mutagenesis experiments within the albino (c) region of mouse chromosome 7 (MMU7). We report here that fit1 mutants are anemic, display numerous peripheral blood defects, and are deficient in early hematopoietic progenitor cell populations. The number of both erythroid and myeloid progenitors, as well as B cells, are reduced. These results implicate fit1 involvement in normal hematopoiesis and suggest that further characterization of the fit1 gene, and the five presumed point mutations of the gene, will lead to an improved understanding of normal hematopoiesis in the mouse.
The Formation of Transferrin Receptor-Positive Sickle Reticulocytes With Intermediate Density Is Not Determined by Fetal Hemoglobin Content
Authors: Franco Robert S. , Thompson Herbert , Palascak Mary , Joiner Clinton H. 
Published: October 2019

AbstractErythrocyte dehydration is an important feature of sickle cell disease, leading to increased sickle hemoglobin polymerization and decreased red blood cell survival. Substantial in vivo dehydration appears to occur in reticulocytes or in an even younger subset of reticulocytes that are positive for transferrin receptor. Previous studies have suggested both sickling-dependent and sickling-independent components of dehydration for these cells. Two types of investigations are reported here. The first series of experiments explored the possibility that fetal hemoglobin (HbF ) content influences the in vivo dehydration of very young, transferrin receptor-positive (T+) cells. These studies confirmed that in most patients the T+ cells in the densest fraction lacked HbF (T+F−). However, T+F− and T+F+ cells appeared to have the same tendency to become moderately dense. The second type of investigation examined moderately dense T+ cells with normalized K+ content and determined the effect of HbF content on KCl cotransport-mediated dehydration in oxygenated incubations. Under these conditions, both T+F− and T+F+ cells had an equal tendency to become more dense by this pathway. Taken together, these studies indicate that at least some young sickle cells become moderately dense due to higher KCl cotransport activity independent of HbF content (and by inference, independent of sickling). However, to become very dense, it appears that further dehydration through a sickling-mediated pathway is required. We suggest that the dehydration of young sickle cells occurs in two steps, with the first dominated by KCl cotransport and the second having an important sickling-dependent component.
Enhanced Levels and Enhanced Clonogenic Capacity of Blood Progenitor Cells Following Administration of Stem Cell Factor Plus Granulocyte Colony-Stimulating Factor to Humans
Authors: Begley C.G. , Basser R. , Mansfield R. , Thomson B. , Parker W.R.L. , Layton J. , To B. , Cebon J. , Sheridan W.P. , Fox R.M. , Green M.D. 
Published: October 2019

AbstractAdministration of hematopoietic growth factors is being used increasingly to obtain populations of blood progenitor/stem cells (PBPC) for clinical transplantation. Here we examined the effect of combining stem cell factor (SCF ) and granulocyte colony-stimulating factor (G-CSF ) versus G-CSF alone in a randomized clinical study involving 62 women with early-stage breast cancer. In the first patient cohorts, escalating doses of SCF were administered for 7 days with concurrent G-CSF administration. At baseline, levels of progenitor cells in the bone marrow or blood were comparable in the different patient groups. As with administration of G-CSF alone, the combination of SCF plus G-CSF did not alter the wide variation in levels of PBPC observed between individuals and did not alter the selective nature of PBPC release, with preferential release of day-14 granulocyte-macrophage colony-stimulating factor (GM-CFC) versus day-7 GM-CFC. However, SCF acted to sustain the levels of PBPC after cessation of growth factor treatment; levels of PBPC were elevated 100-fold at later timepoints compared with G-CSF alone. In addition, the maximum levels of PBPC observed were increased approximately fivefold at day 5 of growth-factor administration. The increased levels of PBPC resulted in significantly increased levels of PBPC obtained by leukapheresis. In a subsequent patient cohort, 3-days pretreatment with SCF was introduced and followed by 7 days concurrent SCF plus G-CSF. The 3-days pretreatment with SCF resulted in an earlier wave of PBPC release in response to commencement of G-CSF. In addition, maximum PBPC levels in blood and PBPC yield in leukapheresis products were further increased. Unexpectedly however, SCF pretreatment resulted in progenitor cells with enhanced self-generation potential. Recloning assays documented the ability of approximately 30% of primary granulocyte-macrophage (GM) colonies from control cell populations to generate secondary GM colonies (n = 1,106 primary colonies examined). In contrast approximately 90% of GM colonies from PBPC after SCF pretreatment generated secondary clones and 65% generated secondary colonies. The action of SCF was not explicable in terms of altered SCF, GM-CSF, or G-CSF responsiveness, but SCF pretreatment was associated with maximum serum SCF levels at the time G-CSF was commenced. These results show that PBPC populations mobilized by different growth factor regimens can differ in their functional properties and caution against solely considering number of harvested progenitor cells without regard to their function.
Adhesion and Migration Are Differentially Regulated in Hematopoietic Progenitor Cells by Cytokines and Extracellular Matrix
Authors: Strobel Eva-Susanne , Möbest Dieter , von Kleist Sabine , Dangel Matthias , Ries Stefan , Mertelsmann Roland , Henschler Reinhard 
Published: October 2019

AbstractThe conditions that control the migratory status of hematopoietic progenitor cells on extracellular matrix (ECM) and that decide whether a cell migrates or adheres are incompletely understood. We analyzed the migratory behavior of murine hematopoietic progenitor cells factor-dependent-cell-paterson (FDCP)-mix and purified lin−Sca1+ bone marrow cells on ECM. We found that migration on fibronectin (Fn) or laminin (Lam) becomes dependent on β1-integrins if a surface restraint force is introduced by tilting the ECM-coated culture vessels. Under these conditions, migration specifically occured on Fn and Lam, and was not detected on collagen IV-, hyaluronate-, or bovine serum albumin- coated surfaces. Migration depended on the continuous presence of hematopoietic cytokines interleukin-3 (IL-3), granulocyte colony-stimulating factor (G-CSF ), macrophage-CSF (M-CSF), granulocyte-macrophage-CSF (GM-CSF ), or stem cell factor (SCF), whereas other cytokines, such as IL-8, macrophage inflammatory protein-1α, macrophage-chemotactic and activating factor, and erythropoietin resulted in very little or no migratory response. IL-3 induced migration was synergistically enhanced by other CSFs, but was completely inhibited by addition of transforming growth factor-β1. In contrast to firm local adhesion of previously cytokine depleted progenitors that was rapidly inducible within 1 hour after exposure to cytokines, preincubation on Fn matrix for 4 to 6 hours was required before cytokines could induce migration. A sudden increase of cytokine concentration reversibly inhibited migration and induced a fully adhesive state; this effect could be prolonged by consecutive stimulation with heterologous cytokines. Whereas cytokines activated resting progenitor cells to migrate on ECM, cell migration speed was regulated by Fn concentration. These results indicate that β1-integrin–mediated progenitor cell adhesion and migration are differentially regulated by external stimuli and suggest that this regulation corresponds to different activation states of β1-integrins in hematopoietic progenitor cells.