bluuebunny/crossref_metadata_2025 huggingface dataset:
Specificity of Effector T Lymphocytes in Autologous Graft-Versus-Host Disease: Role of the Major Histocompatibility Complex Class II Invariant Chain Peptide
Authors: Hess Allan D. , Bright Emilie C. , Thoburn Christopher , Vogelsang Georgia B. , Jones Richard J. , Kennedy M. John
Published: October 2019

Administration of the immunosuppressive drug cyclosporine after autologous bone marrow transplantation induces a systemic autoimmune syndrome resembling graft-versus-host disease (GVHD). This syndrome termed autologous GVHD has significant antitumor activity. Associated with autologous GVHD is the development of T lymphocytes that recognize major histocompatibility complex (MHC) class II determinants, including self. The present studies attempted to characterize and define the molecular specificity of the effector T lymphocytes in autologous GVHD induced in patients with metastatic breast cancer. The results suggest that the effector cells associated with human autologous GVHD are CD8+ T lymphocytes expressing the α/β T-cell receptor. Additional studies show that the effector T cells recognize MHC class II antigens in association with a peptide from the invariant chain (CLIP). Pretreatment of autologous lymphoblast target cells with anti-CLIP antibody completely blocked lysis mediated by autologous GVHD effector T cells. On the other hand, force loading this peptide markedly enhanced the susceptibility of the target cells to recognition by the autoreactive T cells. The recognition of the MHC class II CLIP complex may account for the novel specificity of the effector T cells associated with human autologous GVHD. Moreover, identification of the target peptide may allow for the development of novel immunotherapeutic strategies to enhance the antitumor efficacy of autologous GVHD.
Defects of the Mismatch Repair Gene MSH2 Are Implicated in the Development of Murine and Human Lymphoblastic Lymphomas and Are Associated With the Aberrant Expression of Rhombotin-2 (Lmo-2) and Tal-1 (SCL)
Authors: Lowsky Robert , DeCoteau John F. , Reitmair Armin H. , Ichinohasama Ryo , Dong Wei-Feng , Xu Yi , Mak Tak W. , Kadin Marshall E. , Minden Mark D.
Published: October 2019

Mutations in the DNA mismatch repair (MMR) gene hMSH2 underlie a novel pathway of tumorigenesis for some cancers of epithelial origin. Mice deficient in MSH2 are susceptible to lymphomas but defects in this gene have not been identified in human lymphoid tumors. To determine if the lymphomas these mice develop are related to a particular subtype of human lymphoma we evaluated 20 clinically ill homozygous MSH2−/− mice ranging in age from 2 to 13 months. The murine tumors comprised a single histopathologic entity representing the malignant counterpart of precursor thymic T cells and closely resembled human precursor T-cell lymphoblastic lymphoma (LBL). Evaluation of the expression of three T-cell malignancy associated genes showed that Rhombotin-2 (RBTN-2 also known as Lmo-2), TAL-1 (also known as SCL), and HOX-11 were expressed in 100%, 40%, and 0% of the murine tumors, respectively. The MSH2−/− murine model of precursor T-cell LBL was substantiated by the finding of a nearly identical expression profile of RBTN-2, TAL-1, and HOX-11 in 10 well-characterized cases of human LBL. Direct evidence for MSH2 abnormalities in human LBL was established by sequence analysis of exon 13 of hMSH2, which revealed coding region mutations in 2 of 10 cases. Our findings implicate defects in the MMR system with the aberrant expression of T-cell specific proto-oncogenes and define a new pathway of human lymphomagenesis.
Lineage- and Stage-Specific Expression of Runt Box Polypeptides in Primitive and Definitive Hematopoiesis
Authors: Corsetti Maria Teresa , Calabi Franco
Published: October 2019

Translocations involving the human CBFA2 locus have been associated with leukemia. This gene, originally named AML1, is a human homologue of the Drosophila gene runt that controls early events in fly embryogenesis. To clarify the role of mammalian runt products in normal and leukemic hematopoiesis, we have studied their pattern of expression in mouse hematopoietic tissues in the adult and during ontogeny using an anti-runt box antiserum. In the adult bone marrow, we found expression of runt polypeptides in differentiating myeloid cells and in B lymphocytes. Within the erythroid lineage, runt expression is biphasic, clearly present in the erythroblasts of early blood islands and of the fetal liver, but absent in the adult. Biochemical analysis by Western blotting of fetal and adult hematopoietic populations shows several runt isoforms. At least one of them appears to be myeloid specific.
Regulation of Plasminogen Gene Expression by Interleukin-6
Authors: Jenkins G. Ronald , Seiffert Dietmar , Parmer Robert J. , Miles Lindsey A.
Published: October 2019

Plasmin, the primary fibrinolytic enzyme, has a broad substrate spectrum and participates in other biological processes dependent upon proteolytic activity. Consequently, plasmin activity is tightly regulated by plasminogen activators and protease inhibitors. In this study, we examined whether regulation of plasminogen gene expression also might provide a new mechanism for controlling this system. We examined the effects of recombinant human interleukin-6 (rhIL-6), a pleiotropic cytokine, on plasminogen mRNA expression in primary murine hepatocytes and Hep3B human hepatoma cells. In primary hepatocytes, rhIL-6 and hydrocortisone separately increased plasminogen mRNA expression, but hydrocortisone did not markedly enhance the response to rhIL-6. Hep3B hepatoma cells exhibited more modest responses to rhIL-6. We used the polymerase chain reaction to amplify a 1,067-bp fragment of the human plasminogen promoter/5′ flanking region. This fragment was cloned upstream of a luciferase reporter gene. Hep3B cells transiently transfected with this construct provided ∼100-fold higher luciferase activity compared to cells transfected with control plasmids, and luciferase activity was increased ∼4.5-fold when these cells were treated with rhIL-6. Furthermore, mice injected with rhIL-6 exhibited increases in hepatic plasminogen mRNA. Circulating plasminogen levels were significantly higher in the mice injected with rhIL-6 compared to mice injected with saline. Mice injected with lipopolysaccharide (an inducer of IL-6 in vivo) also showed increased hepatic plasminogen mRNA. Thus, plasminogen gene expression can be modulated by rhIL-6, suggesting a new mechanism for regulating biological systems that use plasmin.
A Molecular Model for the Triplicated A Domains of Human Factor VIII Based on the Crystal Structure of Human Ceruloplasmin
Authors: Pemberton S. , Lindley P. , Zaitsev V. , Card G. , Tuddenham E.G.D. , Kemball-Cook G.
Published: October 2019

The hemophilia A mutation database lists more than 160 missense mutations: each represents a molecular defect in the FVIII molecule, resulting in the X-linked bleeding disorder hemophilia A with a clinical presentation varying from mild to severe. Without a three-dimensional FVIII structure it is in most cases impossible to explain biological dysfunction in terms of the underlying molecular pathology. However, recently the crystal structure of the homologous human plasma copper-binding protein ceruloplasmin (hCp) has been solved, and the A domains of FVIII share approximately 34% sequence identity with hCp. This advance has enabled the building of a molecular model of the A domains of FVIII based on the sequence identity between the two proteins. The model allows exploration of predictions regarding the general features of the FVIII molecule, such as the binding-sites for factor IXa and activated protein C; it has also allowed the mapping of more than 30 selected mutations with known phenotype from the database, and the prediction of hypothetical links to dysfunction in all but a few cases. A computer-generated molecular model such as that reported here cannot substitute for a crystal structure. However, until such a structure for FVIII becomes available, the model represents a significant advance in modeling FVIII; it should prove a useful tool for exploiting the increasing amount of information in the hemophilia A mutation database, and for selecting appropriate targets for investigation of the structure-function relationships via mutagenesis and expression in vitro.
Apoptotic Vascular Endothelial Cells Become Procoagulant
Authors: Bombeli Thomas , Karsan Aly , Tait Jonathan F. , Harlan John M.
Published: October 2019

Whereas unperturbed endothelial cells provide potent anticoagulant properties, exposure to inflammatory and atherogenic stimuli can rapidly lead to a procoagulant behavior. Because recent studies provide evidence that apoptosis of vascular cells may occur under conditions such as atherosclerosis and inflammation, we investigated whether apoptotic endothelial cells may contribute to the development of a prothrombotic state. In this report, it is shown that both adherent and detached apoptotic human umbilical vein endothelial cells (HUVECs) become procoagulant. Apoptosis was induced by staurosporine, a nonspecific protein kinase inhibitor, or by culture in suspension with serum deprivation. Both methods resulted in similar findings. As assessed by flow cytometric determination of annexin V binding, HUVECs undergoing cell death exhibited typically a more rapid exposure of membrane phosphatidylserine (PS) than DNA fragmentation. Depending on the stage of apoptosis, this redistribution of phospholipids was found to induce an increase of the activity of the intrinsic tenase complex by 25% to 60%. Although apoptotic cells did not show antigenic or functional tissue factor (TF ) activity, when preactivated with lipopolysaccharide, TF procoagulant activity increased by 50% to 70%. At 8 hours after apoptosis induction, antigenic thrombomodulin, heparan sulfates, and TF pathway inhibitor decreased by about 83%, 80%, and 59%, respectively. The functional activity of these components was reduced by about 36%, 52%, and 39%, respectively. Moreover, the presence of apoptotic HUVECs led to a significant increase of thrombin formation in recalcified citrated plasma. In conclusion, apoptotic HUVECs, either adherent or in suspension, become procoagulant by increased expression of PS and the loss of anticoagulant membrane components.
Adoptive Transfer of Anti-CD3–Activated CD4+ T Cells Plus Cyclophosphamide and Liposome-Encapsulated Interleukin-2 Cure Murine MC-38 and 3LL Tumors and Establish Tumor-Specific Immunity
Authors: Saxton Mark L. , Longo Dan L. , Wetzel Holly E. , Tribble Henry , Alvord W. Gregory , Kwak Larry W. , Leonard Arnold S. , Ullmann Claudio Dansky , Curti Brendan D. , Ochoa Augusto C.
Published: October 2019

The infusion of anti-CD3–activated murine T cells plus interleukin-2 (IL-2) exerts antitumor effects against several tumors in murine immunotherapy models. This study compares the therapeutic efficacy of anti-CD3–activated CD4+ or CD8+ T-cell subsets, when given with cyclophosphamide (Cy) and liposome-encapsulated IL-2 (L-IL2) in a murine model. C57BL/6 mice bearing subcutaneous (SC) MC-38 colon adenocarcinoma, 3LL Lewis lung carcinoma, or 38C13 lymphoma for 7 to 14 days were pretreated with low-dose intraperitoneal (IP) Cy before intravenous (IV) injection of anti-CD3–activated T cells or T-cell subsets. Cell administration was followed by IP administration of L-IL2 for 5 days. Mice receiving activated CD4+ T cells showed significantly reduced tumor growth or complete remissions with prolonged disease-free survival in MC-38, 3LL, and 38C13. The timing of Cy doses in relation to adoptive transfer was critical in obtaining the optimal antitumor effect by CD4+ cells. Injecting Cy 4 days before the infusion of CD4+ cells greatly enhanced the antitumor effect of the CD4+ cells and improved survival of the mice compared with other Cy regimens. C57BL/6 mice cured of MC-38 after treatment with CD4+ T cells developed tumor-type immunologic memory as demonstrated by their ability to reject rechallenges with MC-38, but not 3LL. Similarly, mice cured of 3LL tumors rejected rechallenges of 3LL, but not MC-38. The immunologic memory could be transferred with an IV injection of splenocytes from mice cured of MC-38 or 3LL. No cytotoxic T-lymphocyte activity was detected in T cells or T-cell subsets from mice cured of MC-38 or 3LL. Increased IL-2 and interferon-γ (IFN-γ) production was observed from CD4+ subsets in cured animals when stimulated in vitro with the original tumor, but not with an unrelated syngeneic tumor. These results suggest that tumor-specific immunity can be achieved in vivo with anti-CD3–stimulated CD4+ T cells in this cellular therapy model.
Decrease of Very Late Activation Antigen-4 and CD36 on Reticulocytes in Sickle Cell Patients Treated With Hydroxyurea
Authors: Styles Lori A. , Lubin Bertram , Vichinsky Elliott , Lawrence Sandy , Hua Mandy , Test Samuel , Kuypers Frans
Published: October 2019

Sickle cell disease (SCD) is characterized by repeated vaso-occlusive events, which result in substantial morbidity. Abnormal adhesion of sickle red blood cells (RBC) to the vascular endothelium is postulated to play a role in the pathogenesis of vaso-occlusion. Two adhesion receptors, very late activation antigen-4 (VLA-4) and CD36, are found in unusually high numbers on sickle cell reticulocytes and do mediate adhesion of sickle RBC to endothelium. Hydroxyurea (HU) therapy results in fewer vaso-occlusive episodes, and we postulated that HU-related modulation of VLA-4 and CD36 receptors may contribute to its clinical benefit. Using flow cytometry, eight patients were followed from the onset of HU treatment through a mean treatment length of 200 ± 49 days. Mean corpuscular volume and percent fetal hemoglobin (Hb F ) increased from 87% ± 6% to 98% ± 9% and 6.6% ± 3.9% to 12.7% ± 5.6%, respectively. The percentage of reticulocytes expressing VLA-4 decreased from 29.0% ± 5.9% to 14.9% ± 2.3% (P = .0003). Two thirds of the total decrease in VLA-4 expression occurred after 10 weeks of HU and plateaued by 20 weeks. Changes in VLA-4 expression occurred before substantial increases in Hb F. The percentage of reticulocytes expressing CD36 decreased from 55.3% ± 6.4% to 42.6% (P = .0046). Changes in adhesion receptor expression were not caused by a decrease in reticulocytosis with HU therapy. This report is the first to associate a decrease in adhesion receptor expression with a therapy known to reduce the clinical severity of SCD.
The Characterization, Molecular Cloning, and Expression of a Novel Hematopoietic Cell Antigen From CD34+ Human Bone Marrow Cells
Authors: Uchida Nobuko , Yang Zhi , Combs Jesse , Pourquié Olivier , Nguyen Megan , Ramanathan Rajeev , Fu Joan , Welply Annemarie , Chen Shirley , Weddell Greg , Sharma Arun K. , Leiby Kevin R. , Karagogeos Domna , Hill Beth , Humeau Laurent , Stallcup William B. , Hoffman Ron , Tsukamoto Ann S. , Gearing David P. , Péault Bruno
Published: October 2019

The adhesion molecule BEN/SC1/DM-GRASP (BEN) is a marker in the developing chicken nervous system that is also expressed on the surface of embryonic and adult hematopoietic cells such as immature thymocytes, myeloid progenitors, and erythroid progenitors. F84.1 and KG-CAM, two monoclonal antibodies to rat neuronal glycoproteins with similarity to BEN, cross-react with an antigen on rat hematopoietic progenitors, but F84.1 only also recognizes human blood cell progenitors. We have defined the antigen recognized by F84.1 as the hematopoietic cell antigen (HCA). HCA expression was detected on 40% to 70% of CD34+ fetal and adult bone marrow cells and mobilized peripheral blood cells. Precursor cell activity for long-term in vitro bone marrow cell culture was confined to the subset of CD34+ cells that coexpress HCA. HCA is expressed by the most primitive subsets of CD34+ cells, including all rhodamine 123lo, Thy-1+, and CD38−/lo CD34+ adult bone marrow cells. HCA was also detected on myeloid progenitors but not on early B-cell progenitors. We also describe here the cloning and characterization of cDNAs encoding two variants of the human HCA antigen (huHCA-1 and huHCA-2) and of a cDNA clone encoding rat HCA (raHCA). The deduced amino acid sequences of huHCA and raHCA are homologous to that of chicken BEN. Recombinant proteins produced from either human or rat HCA cDNAs were recognized by F84.1, whereas rat HCA but not human HCA was recognized by antirat KG-CAM. Expression of either form of huHCA in CHO cells conferred homophilic adhesion that could be competed with soluble recombinant huHCA-Fc. The molecular cloning of HCA and the availability of recombinant HCA should permit further evaluation of its role in human and rodent hematopoiesis.
Selective Expression of the Receptor Tyrosine Kinase, HTK, on Human Erythroid Progenitor Cells
Authors: Inada Tomohisa , Iwama Atsushi , Sakano Seiji , Ohno Mitsuharu , Sawada Ken-ichi , Suda Toshio
Published: October 2019

HTK is a receptor tyrosine kinase of the Eph family. To characterize the involvement of HTK in hematopoiesis, we generated monoclonal antibodies against HTK and investigated its expression on human bone marrow cells. About 5% of the bone marrow cells were HTK+, which were also c-Kit+, CD34low, and glycophorin A−/low. Assays of progenitors showed that HTK+c-Kit+ cells consisted exclusively of erythroid progenitors, whereas HTK−c-Kit+ cells contained progenitors of granulocytes and macrophages as well as those of erythroid cells. Most of the HTK+ erythroid progenitors were stem cell factor-dependent for proliferation, indicating that they represent mainly erythroid burst-forming units (BFU-E). During the erythroid differentiation of cultured peripheral CD34+ cells, HTK expression was upregulated on immature erythroid cells that corresponded to BFU-E and erythroid colony-forming units and downregulated on erythroblasts with high levels of glycophorin expression. These findings suggest that HTK is selectively expressed on the restricted stage of erythroid progenitors, particularly BFU-E, and that HTK is the first marker antigen that allows the purification of erythroid progenitors. Furthermore, HTKL, the ligand for HTK, was expressed in the bone marrow stromal cells. Our findings provide a novel regulatory system of erythropoiesis mediated by the HTKL-HTK signaling pathway.
Oligoclonal CD4+CD57+ T-Cell Expansions Contribute to the Imbalanced T-Cell Receptor Repertoire of Rheumatoid Arthritis Patients
Authors: Imberti Luisa , Sottini Alessandra , Signorini Simona , Gorla Roberto , Primi Daniele
Published: October 2019

A peculiar feature of rheumatoid arthritis patients is that they carry clonally expanded CD4+ and CD8+ cells in the peripheral blood. While the distortion of the repertoire of CD8+ cells has been ascribed to the increase of CD8+CD57+ large granular lymphocytes, often detected in these patients, the mechanism responsible for the clonal expansion of CD4+ cells remains unexplained. Here, we report that CD4+CD57+ cells, that in healthy individuals represent a small subset of peripheral CD4+ lymphocytes, are significantly expanded in the peripheral blood of a considerable percentage of rheumatoid arthritis patients. Furthermore, the expansion of these lymphocytes appears to correlate with the presence of rheumatoid factor. The molecular analysis of the T-cell receptor variable beta segments expressed by the CD4+CD57+ cells enriched in rheumatoid arthritis patients showed that they use restricted repertoires, that partially overlap with those of their CD4−CD57+ counterpart. The structural feature of the receptor ligand expressed by these cells revealed that their expansion is most likely mediated by strong antigenic pressures. However, since we also found that CD4+CD57+ and CD4−CD57+ cells can share the same clonal specificity, it is likely that their selection is not mediated by conventional major histocompatibility complex restricted mechanisms. Thus, while our data demonstrate that CD4+CD57+ cells play an important role in establishing the imbalance of the CD4+ cell repertoire observed in rheumatoid arthritis patients, they also suggest that these cells have common features with mouse CD4+CD8−NK1.1+/T cells.
Immunostimulatory Oligodeoxynucleotides Containing CpG Motifs Enhance the Efficacy of Monoclonal Antibody Therapy of Lymphoma
Authors: Wooldridge James E. , Ballas Zuhair , Krieg Arthur M. , Weiner George J.
Published: October 2019

Bacterial DNA and synthetic oligodeoxynucleotides containing the CpG motif (CpG ODN) can activate various immune cell subsets, including natural killer cells and macrophages. We evaluated whether the combination of CpG ODN and antitumor monoclonal antibody is effective at preventing tumor growth in an immunocompetent murine lymphoma model. CpG ODN–activated murine splenocytes induced lysis of tumor targets more effectively than unactivated splenocytes. These effector cells were also superior to unactivated splenocytes or cells activated with a control methylated ODN at inducing antibody-mediated lysis of 38C13 murine lymphoma cells. In vivo, CpG ODN alone had no effect on survival of mice inoculated with 38C13 cells. However, a single injection of CpG ODN enhanced the antitumor response to antitumor monoclonal antibody therapy. Ninety percent of mice treated with monoclonal antibody alone developed tumor compared with 20% of mice treated with antibody and CpG ODN. These antitumor effects were less pronounced when treatment consisted of an identical ODN containing methylated CpG dinucleotides. A single dose of CpG ODN appeared to be as effective as multiple doses of interleukin-2 at inhibiting tumor growth when combined with antitumor monoclonal antibody. We conclude that immunostimulatory CpG ODN can enhance antibody dependent cellular cytotoxicity and warrant further evaluation as potential immunotherapeutic reagents in cancer.
Donor Leukocyte Infusion for Leukemic Relapse After Allogeneic Marrow Transplantation: Lack of Residual Donor Hematopoiesis Predicts Aplasia
Authors: Keil Felix , Haas Oskar A. , Fritsch Gerhard , Kalhs Peter , Lechner Klaus , Mannhalter Christine , Reiter Elisabeth , Niederwieser Dietger , Hoecker Paul , Greinix Hildegard T.
Published: October 2019

We assessed the chimerism of CD34+ bone marrow cells before donor leukocyte infusion (DLI) on nine occasions in seven patients with leukemic relapse after allogeneic marrow transplantation. The patients suffered from acute lymphoblastic leukemia (n = 1), acute myeloid leukemia (n = 3), and chronic myeloid leukemia (CML; n = 3). Two patients received a second DLI because of disease progression after the first one. The origin of the CD34+ cells was determined by analyzing variable number of tandem repeats with polymerase chain reaction and, in sex-mismatched cases, by fluorescence in situ hybridization. Before DLI CD34+ cells were exclusively of donor origin in four patients. In another patient 41% of CD34+ cells were derived from the donor. No aplasia occurred in these patients after DLI, whereas in the two patients with exclusively recipient hematopoiesis severe aplasia lasting for 5 and 13 weeks necessitated hematopoietic stem cell support. One patient who had only 5% CD34+ donor cells before DLI recovered without stem cell support after 10 days. Two patients in relapse of CML showed a high percentage of BCR-ABL− CD34+ cells of recipient origin before DLI. These BCR-ABL− cells of recipient type did not prevent severe aplasia which indicates that the assessment of BCR-ABL+ hematopoiesis alone is insufficient for predicting aplasia. Our data indicate that in case of sufficient donor hematopoiesis before DLI no persistent aplasia will occur. Thus, evaluation of donor hematopoiesis allows prediction of aplasia after DLI and makes early therapeutic interventions possible.
Synergistic Effect of FLT-3 Ligand on the Granulocyte Colony-Stimulating Factor–Induced Mobilization of Hematopoietic Stem Cells and Progenitor Cells Into Blood in Mice
Authors: Sudo Yoshikazu , Shimazaki Chihiro , Ashihara Eishi , Kikuta Takehisa , Hirai Hideyo , Sumikuma Toshiya , Yamagata Noboru , Goto Hideo , Inaba Tohru , Fujita Naohisa , Nakagawa Masao
Published: October 2019

We have previously shown that FLT-3 ligand (FL) mobilizes murine hematopoietic primitive and committed progenitor cells into blood dose-dependently. Whether FL also acts synergistically with granulocyte colony-stimulating factor (G-CSF ) to induce such mobilization has now been investigated. Five- to 6-week-old C57BL/6J mice were injected subcutaneously with recombinant human G-CSF (250 μg/kg), Chinese hamster ovarian cell-derived FL (20 μg/kg), or both cytokines daily for 5 days. The number of colony-forming cells (CFCs) in peripheral blood increased approximately 2-, 21-, or 480-fold after administration of FL, G-CSF, or the two cytokines together, respectively, for 5 days. The number of CFCs in bone marrow decreased after 3 days but was increased approximately twofold after 5 days of treatment with G-CSF. The number of CFCs in the bone marrow of mice treated with both FL and G-CSF showed a 3.4-fold increase after 3 days and subsequently decreased to below control values. The number of CFCs in spleen was increased 24.2- and 93.7-fold after 5 days of treatment with G-CSF alone or in combination with FL, respectively. The number of colony-forming unit-spleen (CFU-S) (day 12) in peripheral blood was increased 13.2-fold by G-CSF alone and 182-fold by G-CSF and FL used together after 5 days of treatment. Finally, the number of preCFU-S mobilized into peripheral blood was also increased by the administration of FL and G-CSF. These observations show that FL synergistically enhances the G-CSF–induced mobilization of hematopoietic stem cells and progenitor cells into blood in mice, and that this combination of growth factors may prove useful for obtaining such cells in humans for transplantation.
Focal Adhesion Kinase Upregulated by Granulocyte-Macrophage Colony-Stimulating Factor But Not by Interleukin-3 in Differentiating Myeloid Cells
Authors: Kume Akihiro , Nishiura Hiroshi , Suda Junko , Suda Toshio
Published: October 2019

The involvement of focal adhesion kinase (FAK) in myeloid differentiation was investigated in primary murine bone marrow (BM) cells. In unstimulated BM, FAK mRNA was detected in myeloid and lymphoid cells, but not in erythroid precursors. When the BM cells were incubated with granulocyte-macrophage colony-stimulating factor (GM-CSF ) or interleukin-3 (IL-3), FAK expression showed a remarkable difference depending on the cytokine. Although FAK was upregulated in the cells stimulated by GM-CSF (GM-treated cells), the kinase was barely detectable in the cells cultured with IL-3 (IL-3–treated cells). Morphology and flow cytometry analysis showed GM-CSF promoted the growth and differentiation of monocyte/macrophage lineage stronger than IL-3. In addition, motility of the cytokine-differentiated cells showed an overt distinction between the cultures, which was closely correlated with FAK expression. After 7 days of stimulation, GM-treated cells showed active migration and chemoattractant-induced morphologic polarization. In contrast, IL-3–treated cells showed minimal migration and polarization. These results suggest an important role of GM-CSF in the terminal differentiation of monocytes/macrophages, and possible involvement of FAK in functional maturity of this lineage.
Case Reports: Familial Hodgkin's Disease
Authors: SCHIER W. WILSON
Published: October 2019

The multiple occurrence of Hodgkin’s and allied diseases in each of two different families is reported.

The infrequent appearance of Hodgkin’s disease in the general population suggests that the familial incidence may be more than fortuitous.

The significance of familial Hodgkin’s disease is briefly discussed in the light of current thought concerning the etiology of Hodgkin’s disease.
The Erythropoietic Effect of Urine and Plasma of Repeatedly Bled Rabbits
Authors: HODGSON G. , TOHÁ J.
Published: October 2019

1. The reticulocyte response of rabbits bled in a standard manner is significantly greater after the injection of anemic rabbit plasma, than after the injection of normal rabbit plasma. The effect of anemic rabbit plasma disappears after the exposure to 100 per cent oxygen, but not after exposure to air. The activity of anemic rabbit plasma is not due to the presence of high concentrations of vitamin B12 or folic acid.

2. The recovery of hemoglobin concentration after a standard hemorrhage is described by a first order exponential equation. The speed of recovery is characterized by the time constant of regeneration. The velocity of hemoglobin regeneration, after a standard hemorrhage in rabbits injected with plasma of anemic animals, is greater than that of controls injected with normal rabbit plasma. Anemic rabbit plasma loses its erythropoietic effect after boiling at pH 5.5 for 15 minutes.

3. The repeated injection of anemic rabbit plasma in normal rabbits produces a significant rise of reticulocytes, erythrocytes, and hemoglobin concentration.

4. The injection of urine of anemic animals increases the speed of hemoglobin regeneration after a standard hemorrhage in rabbits. The activity of the urine of anemic rabbits is lost by boiling at pH 9.0 for 5 minutes, but not after the same treatment at pH 5.5. Urine of anemic animals loses its erythropoietic action after 16 hours dialysis.

5. It is concluded that the plasma and urine of animal with increased erythropoiesis contains a factor or factors (erythropoietins) which are probably the humoral mediators of the primary erythropoietic stimulus.
The Lysis of Intravascular Thrombi in Rabbits with Human Plasmin (Fibrinolysin)
Authors: GROSSI CARLO E. , CLIFFTON EUGENE E. , CANNAMELA DOLLY A.
Published: October 2019

1. Thrombi were produced in the marginal vein of the ear of rabbits with the use of either thrombin or sodium morrhuate. Sodium morrhuate was found to be most dependable.

2. Plasmin given intravenously with an average dose of 10 to 15 ml. per Kg. was effective in producing a complete intravascular lysis in all instances where the thrombus was less than 24 hours old, and a partial or total lysis in the great majority of animals with thrombi over 24 hours old. This lysis took place within 1 to 2 hours after the injection of plasmin.

3. The intravenous use of plasmin is shown in rabbits to be nonlethal and nontoxic.
Hematologic Responses and Concenrtation of Vitamin B12 in Serum and Urine following Oral Administration of Vitamin B12 without Intrinsic Factor
Authors: ROSS G. I. M. , MOLLIN D. L. , COX E. V. , UNGLEY C. C.
Published: October 2019

Vitamin B12 in doses of 500 to 3000 µg. was given by mouth or by rectum to patients with megaloblastic anemia due to B12 deficiency and to normal subjects. Its absorption was studied by following the hematologic response and/or the changes in serum and urinary B12 concentrations by microbiologic assay with Euglena gracilis var. bacillaris.

After oral doses of 1000 to 3000 µg. of B12 there was a definite but variable hematologic response in the thirty-five patients. The serum B12 concentrations, measured in eighteen of these patients, increased in the first twenty-four hours after the dose. The urinary excretion of B12 increased in fifteen out of sixteen patients, but the average amount excreted was only one quarter of that excreted after intramuscular injections producing comparable hematologic responses. The lower urinary excretion after the oral doses was associated with a lower total and uncombined B12 concentration in the serum. In the majority of patients serum B12 concentrations and urinary B12 excretion reached a maximum in the first six hours after the dose.

After oral doses of 3000 µg. there was a rise in serum and urinary B12 in three out of four normal subjects.

After doses of 500 µg., there was an increase in the serum B12 concentrations of two normal subjects and of one out of two patients with megaloblastic anemia, but there was no change in urinary B12 excretion.

After rectal doses of 1000 or 3000 µg. there was evidence of absorption of the vitamin in seven out of eight patients with pernicious anemia.
Analytical Review: The Mechanism of Glucocorticoid Eosinopenia
Authors: ESSELLIER A. F. , JEANNERET R. L. , MORANDI L.
Published: October 2019

This paper deals with basic problems of glucocorticoid eosinopenia and of regulations within the eosinophile cell system.

The effects of glucocorticoids on single regulatory factors of this system, such as eosinotactic stimuli, eosinopoiesis, life span and rate of removal of eosinophils, are discussed in detail. Eosinophilopoiesis is not influenced by glucocorticoids. This finding, added to the observation that blocking the reticuloendothelial system suppresses the glucocorticoid eosinopenia and that these compounds have no eosinolytic properties in vitro, speaks against the contention that the eosinopenic effect of the glucocorticoids is actually due to a direct lytic effect on the eosinophils. A distributional shift of eosinophils in vascular sectors functioning as blood depots, or a migration into tissues can be abandoned as explanations of glucocorticoid eosinopenia. The hypothesis of a fixation at sites of physiologic demand is certainly not a valid explanation of glucocorticoid eosinopenia, since glucocorticoids suppress the eosinotactic stimuli. Blocking experiments of the RES show that the rate of disappearance of the eosinophils is a resultant of the functional condition of the RES and of the sum of the eosinopenic stimuli.

On the basis of our investigations we arrive at the conclusion that glucocorticoid eosinopenia depends upon an inhibition of the release of mature eosinophils from the bone marrow and upon an increased destruction of eosinophils in the reticuloendothelial system. The level of the original eosinophilia, that is to say, the percentage of young forms with a long life expectancy, determines the degree of eosinopenia attained by a single application of glucocorticoid and the time lapse before aneosinophilia develops under continuous glucocorticoid administration.
Structural Changes in the Platelets as Observed by Electron Microscopy
Authors: BRAUNSTEINER H. , FELLINGER K. , PAKESCH F.
Published: October 2019

Deficient pseudopod formation and spreading of platelets on Formvar films were encountered in cases of chronic idiopathic thrombocytopenic purpura and thrombocytoasthenias. The same defect together with agglutination was induced in isolated normal platelets by a platelet agglutinin from the serum of one case of chronic ITP. The possible significance of these results is briefly discussed.
Idiopathic Immunoneutropenia
Authors: MÅRTENSSON JOHAN , VIKBLADH INGA
Published: October 2019

A case report is presented of a woman, aged 49, with persistent neutropenia, increased erythrocyte sedimentation rate, a pathologic serum protein fraction migrating at the same rate as γ-globulin representing about 30 per cent of the total protein, and with undue fatigue as the only symptom.

Normal leukocytes, especially neutrophils, were agglutinated by a factor contained in the patient’s serum and bound to the pathologic γ-globulin fraction and active even in a dilution of at least 1:120.

The neutropenia may be explained by the formation, in the patient, of antibodies intravascularly agglutinating her own neutrophile leukocytes, which were then eliminated from the circulating blood. The appearance of the bone marrow did not argue against such an assumption.

It has not been possible to demonstrate any definite causal factor of the disease. As far as the neutrophile leukocytes are concerned, the disease is analogous with idiopathic immunohemolytic anemia and idiopathic immunothrombocytopenia.

No leukocyte agglutination could be demonstrated in sera from two other patients with granulocytopenia or in sera from a number of patients with various diseases and associated increased γ-globulin.
Mediterranean Anemia in a Negro Complicated by Pernicious Anemia of Pregnancy
Authors: GOLDBERG MELVIN A. , SCHWARTZ STEVEN O.
Published: October 2019

1. A case is presented of a Negro woman with Mediterranean anemia who developed megaloblastic anemia of pregnancy in each of three pregnancies.

2. The possibility that Mediterranean anemia predisposes to the development of pernicious anemia of pregnancy is discussed.

3. With megaloblastic erythropoiesis, anemia became more severe, but the characteristic red cell hypochromia of Mediterranean disease was masked.

4. Purified liver extract failed to protect against the development of megaloblastic anemia of pregnancy but folic acid produced a satisfactory therapeutic response.
The Effects of Cortisone on the Development of Spontaneous Leukemia in Mice and on Its Induction by Irradiation
Authors: UPTON A. C. , FURTH J.
Published: October 2019

The development of spontaneous lymphoma in a high leukemia strain of mice was inhibited by administration of cortisone, although the hormone-induced atrophy of the thymus and other lymphoid tissues was only transient.

Inhibition of lymphoma formation resulted from only three successive daily injections of cortisone but was greater if the cortisone administration was prolonged for three months.

A high incidence of myeloid leukemia was induced in a low-leukemia strain of mice by a single exposure to 350 r of x-radiation.

Cortisone was without effect on the induction of myeloid leukemia.

Cortisone administered preirradiation increased, and postirradiation decreased, the incidence of radiation-induced lymphomas in male mice.
Leukoagglutinins
Authors: DAUSSET J. , NENNA A. , BRECY H.
Published: October 2019

1. A simple test for the detection of leukoagglutinating properties of serum is described.

2. A thermolabile serum system which inhibits the leukoagglutination was observed.

3. Sera of about two thousand normal subjects and six hundred patients with various diseases were tested. The sera of nineteen of the patients gave positive reactions.

4. The serologic and physicochemical properties of the leukoagglutinating substance were studied, and evidence is presented in favor of its being an antibody.

5. The role of the leukoagglutinating substance in the development of leukopenia is discussed, as well as the probable type of antibody responsible for the leukoagglutination reaction.
Evidence for a Humoral Regulation of Erythropoiesis
Authors: STOHLMAN FREDERICK , RATH CHARLES E. , ROSE JOHN C.
Published: October 2019

Data obtained from a patient with polycythemia secondary to a patent ductus arteriosus with reverse flow, have been presented. Oxygen saturation and tension of brachial arterial blood and sternal bone marrow were within normal limits, while those of the iliac marrow and femoral arterial blood were reduced.

The presence of polycythemia under such circumstances is thought to exclude a primary cerebral or pituitary mechanism. Similarly, the presence of hypercellularity and normoblastic hyperplasia in normally oxygenated areas of the bone marrow argue against a direct effect of lowered O2 content or pO2 on germinal erythroid cells.

These observations are compatible with a humoral regulation of erythropoiesis in the hypoxic state.
Congenital Afibrinogenemia
Authors: ALEXANDER BENJAMIN , GOLDSTEIN ROBERT , RICH LESTER , LE BOLLOC'H ANNE G. , DIAMOND LOUIS K. , BORGES WAYNE
Published: October 2019

In a study of three subjects with incoagulable blood due to congenital afibrinogenemia, information is presented regarding some basic aspects of coagulation independent, of the fibrinogen-fibrin conversion mechanism. The following facts have been established: (1) Shortly after the blood is exposed to glass, the earliest detectable changes are morphologic alteration in the platelets and their progressive agglutination and lysis. (2) Almost in parallel, plasma antihemophilic activity, originally normal, declines rapidly and practically disappears. (3) Also within minutes, SPCA evolves from its precursor in a normal manner. (4) Acglobulin is rapidly consumed. (5) These changes are well along before detectable amounts of prothrombin have disappeared. (6) Prothrombin consumption proceeds at normal velocity, or slightly faster than normal. (7) Thrombin addition to afibrinogenemic plasma induces platelet agglutination, but this does not occur when thrombin is mixed with platelets alone. (8) The "natural" antithrombic activity of afibrinogenemic plasma is normal. (9) The heparin co-factor of antithrombic activity was demonstrable. (10) Quantitative data were obtained relating the one-stage prothrombin time with the fibrinogen concentration.

The theoretic and practical implications of the observations are discussed.
Results of High-Dose Therapy and Autologous Bone Marrow/Stem Cell Transplantation During Remission in Poor-Risk Intermediate- and High-Grade Lymphoma: International Index High and High-Intermediate Risk Group
Authors: Nademanee Auayporn , Molina Arturo , O'Donnell Margaret R. , Dagis Andrew , Snyder David S. , Parker Pablo , Stein Anthony , Smith Eileen , Planás Ina , Kashyap Ashwin , Spielberger Ricardo , Fung Henry , Wong K.K. , Somlo George , Margolin Kim , Chow Warren , Sniecinski Irena , Vora Nayana , Blume Karl G. , Niland Joyce , Forman Stephen J.
Published: October 2019

We have conducted a pilot study to investigate the role of high-dose therapy and autologous bone marrow/stem cell transplantation (ASCT) during first complete or partial remission in 52 patients with poor-risk aggressive lymphoma. There were 42 patients with intermediate-grade or immunoblastic lymphoma who were considered to be high (60%) and high-intermediate risk (40%) groups at diagnosis based on the age-adjusted International Prognostic Index (IPI) and 10 patients with high-grade, SNCCL (small non-cleaved cell, Burkitt's, and non-Burkitt's), who at presentation had poor-risk features defined as elevated serum lactate dehydrogenase level, stage IV, and bulky mass ≥10 cm. The median age was 34 years (range, 16 to 56 years). Thirty-nine were transplanted in first complete remission and 13 in first partial remission after conventional therapy. Conditioning regimens consisted of total body irradiation (TBI) administered as a single fraction 750 cGy in 3 patients and in fractionated doses for a total of 1,200 cGy in 44 patients, in combination with 60 mg/kg etoposide and 100 mg/kg cyclophosphamide. Five patients with prior radiotherapy received 450 mg/m2 carmustine instead of TBI. Stem cell sources were either bone marrow and/or peripheral blood. No in vitro purging was used. All patients engrafted. Two SNCCL patients died of venoocclusive disease at 25 days and acute leukemia at 27 months posttransplantation. There were six relapses at 1.5 to 12.8 months posttransplantation. At a median follow-up of 44 months (range, 1 to 113 months), the estimated 3-year overall survival (OS) and disease-free survival (DFS) for all patients was 84% (95% confidence interval [CI], 70% to 92%) and 82% (95% CI, 68% to 91%), respectively. In the subset of patients with intermediate-grade and immunoblastic lymphoma, the 3-year DFS was 89% (95% CI, 74% to 96%) for all patients, 87% (95% CI, 67% to 96%) for high-risk patients, and 92% (95 CI, 61% to 99%) for high-intermediate risk patients. The 3-year OS and DFS for SNCCL patients were identical at 60% (95% CI, 30% to 84%). These results suggest that high-dose therapy and ASCT during first remission may improve the survival and prognosis of patients with poor-risk intermediate- and high-grade lymphoma. A prospective randomized study comparing high-dose therapy and ASCT with conventional chemotherapy in IPI high-risk patients with aggressive non-Hodgkin's lymphoma should be undertaken.
Recombinant Human Interleukin-11 Directly Promotes Megakaryocytopoiesis In Vitro
Authors: Weich Nadine S. , Wang Anlai , Fitzgerald Michael , Neben Tamlyn Yee , Donaldson Debra , Giannotti JoAnn , Yetz-Aldape Joanne , Leven Robert M. , Turner Katherine J.
Published: October 2019

We have investigated the mechanism of action of the thrombopoietic cytokine, recombinant human interleukin-11 (rhIL-11), on megakaryocytopoiesis in vitro. We have shown that rhIL-11–induced murine and human megakaryocytopoiesis are not mediated by thrombopoietin (Tpo). Murine megakaryocytes (MKs) were produced from bone marrow (BM) mononuclear cells cultured with rhIL-11, IL-3, and a combination of the two cytokines. Conditioned media (CM) were collected and assayed for the presence of biologically active Tpo. Tpo activity was not detected in any of the CMs tested. Next, human BM CD34+ cells were cultured in serum-free fibrin clot medium with rhIL-11, IL-3, or rhIL-11 plus IL-3 and an antibody that neutralizes human Tpo activity. No inhibition of either burst-forming unit-MK– or colony-forming unit-MK–derived colony formation was observed. The antibody did partially inhibit steel factor-induced MK-colony formation, suggesting that the actions of this cytokine are mediated, in part, by Tpo. We determined that MKs can be direct targets of rhIL-11 by showing the expression of functional IL-11 receptor on these cells. Total RNA was prepared from cultured human BM CD41+CD14− cells (MKs) and IL-11 receptor α chain mRNA was detected in the MKs by reverse transcription-polymerase chain reaction. Analysis of single-sorted CD41+CD14− cells confirmed that the observed IL-11 receptor expression was not due to contaminating CD41− cells in the pool. The presence of rhIL-11 receptor α chain protein in the cells was established by Western blot analysis. After a short exposure of purified BM MKs to rhIL-11, enhanced phosphorylation of both its signal transduction subunit, gp130, and the transcription factor, STAT3 was detected, showing a direct activation of receptor signaling by the cytokine. Consistent with the lack of effect of rhIL-11 on platelets in vivo, IL-11 receptor α chain mRNA and protein were not detected in isolated human platelets. These data indicate that rhIL-11 acts directly on MKs and MK progenitors but not on platelets.
Regulation of Myosin Phosphatase Through Phosphorylation of the Myosin-Binding Subunit in Platelet Activation
Authors: Nakai Keiji , Suzuki Yoshinori , Kihira Hisakazu , Wada Hideo , Fujioka Masanori , Ito Masaaki , Nakano Takeshi , Kaibuchi Kozo , Shiku Hiroshi , Nishikawa Masakatsu
Published: October 2019

Human platelets were found to contain myosin phosphatase consisting of a 38-kD catalytic subunit of protein phosphatase type 1δ, a 130-kD myosin-binding subunit (MBS) and a 20-kD subunit, all of which cross-reacted with antibodies against these subunits of smooth muscle myosin phosphatase. Anti-MBS antibody coimmunoprecipitated RhoA and Rho-kinase of human platelets. Platelets MBS is a substrate for Rho-kinase and phosphorylation of MBS decreases the activity of myosin phosphatase. Treatment of intact platelets with 9,11-epithio-11,12-methano-thromboxane A2 led to a dramatic increase in phosphorylation of MBS and a significant decrease in the activity of myosin phosphatase. These findings suggest a putative mechanism for agonist-induced regulation of myosin phosphatase activity in platelets.