bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular organization of the cytokine gene cluster, involving the human IL-3, IL-4, IL-5, and GM-CSF genes, on human chromosome 5
Authors: van Leeuwen BH , Martinson ME , Webb GC , Young IG 
Published: October 2019

Abstract
The human genes for the hematopoietic growth factors interleukin-3 (IL- 3), IL-5, and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been mapped to 5q23–31. We present in situ hybridization evidence that the human IL-4 gene is located at 5q23.3–31.2, suggesting that the four cytokine genes may be closely linked. We used pulsed-field gel electrophoresis to prepare subchromosomal restriction maps surrounding these genes to define this possible linkage more precisely. The IL-4 and IL-5 genes are tightly linked, being 90 to 240 kilobases (kb) apart, as has been shown for the IL-3 and GM-CSF genes, which are only 9 kb apart. Possible overlap of the map containing the IL-4 and IL-5 genes with restriction sites 5′ to the IL-3 gene suggests that the four cytokine genes may be localized within 500 kb of each other. The endothelial cell growth factor gene (ECGF), which has also been localized to the 5q31 region, did not appear to be close to the cytokine genes. Linkage of the IL-3, IL-4, IL-5, and GM-CSF genes has important implications in the evolutionary origin and regulation of expression of these genes. The four cytokine genes are located in the region of the long arm of chromosome 5, which is deleted in the 5q- anomaly. The present study provides a basis for further investigations of this disorder.
Effects of human interleukin-3 on granulocytic colony-forming cells in human bone marrow
Authors: Bot FJ , van Eijk L , Schipper P , Lowenberg B 
Published: October 2019

Abstract
Human multilineage colony-stimulating factor (multi-CSF)/interleukin-3 (IL-3) induces colony formation from CFU-GEMM, BFU-E, and CFU-Eo when applied to in vitro cultures of highly enriched hematopoietic progenitor cells. No granulocytic colonies are formed in response to IL- 3. However, with appropriate assays, we demonstrate that IL-3 increases the size of G-CSF-induced granulocytic colonies; these colonies contain greater proportions of immature cells as compared with colonies stimulated by G-CSF alone. Furthermore, IL-3 promotes the survival of CFU-G in vitro, whereas in cultures not supplemented with IL-3, CFU-G extinguish within seven days. We conclude that IL-3, although it does not stimulate granulocytic colony formation by itself, regulates the survival and proliferative rate of granulocytic progenitors.
Factor-independent erythropoietic progenitor cells in leukemia induced by the myeloproliferative leukemia virus
Authors: Wendling F , Penciolelli JF , Charon M , Tambourin P 
Published: October 2019

Abstract
The myeloproliferative leukemia virus (MPLV), a novel murine retroviral complex that does not transform fibroblasts, has been shown to cause an acute leukemia in adult mice accompanied by a progressive polycythemia. The present study demonstrates that, on in vivo inoculation, MPLV induces a rapid suppression of growth factor requirement for in vitro colony formation by both the late and the primitive erythroid progenitor cells. CFU-e-derived erythrocytic colonies developed and differentiated in semi-solid medium without the addition of erythropoietin (Epo). In addition, the formation of CFU-e colonies was not altered by the presence of specific neutralizing Epo antibodies. In the spleen, the CFU-e pool size increased rapidly up to 30-fold. By day 6 postinfection, 100% of these progenitor cells were Epo-independent. The in vivo effects of MPLV-infection on early erythroid progenitor cell compartments were examined in cultures grown for seven days. The concentration of erythroid progenitor cells was twofold elevated in spleen from MPLV-infected mice. As early as day 4 postinfection, 50% of these progenitors produced fully hemoglobinized colonies in serum-free cultures without the addition of interleukin-3 (IL-3) and Epo. Most spontaneous colonies were large and contained up to 10(5) cells per colony. They were composed of either erythroblasts only (16%) or erythroblasts and megakaryocytes (70%); few of them were multipotential (14%). In the marrow, the total number of BFU-e was reduced and only few factor-independent bursts were observed, suggesting a rapid migration of infected progenitors from marrow to spleen. Furthermore, the data show that abnormal erythropoiesis was due to the replication defective MPLV information and was not influenced by the Fv-2 locus.
Interleukin-3 down-regulates its own receptor
Authors: Murthy SC , Sorensen PH , Mui AL , Krystal G 
Published: October 2019

Abstract
To gain insight into the mechanisms involved in regulating murine interleukin-3 (mIL-3) receptor expression, we have examined the effects of mIL-3 and murine granulocyte-macrophage colony-stimulating factor (mGM-CSF) on mIL-3 receptor internalization and re-expression and studied the relationship between mIL-3 cell surface receptor density and growth factor sensitivity. As a source of cells for our studies, we used a B6SUtA clone, B6SUtA1, which grows equally well in mIL-3 or mGM- CSF when supplemented with 20% fetal calf serum (FCS) in RPMI 1640. Intracellular processing studies carried out in the presence and absence of methylamine suggested that mIL-3 is cleaved at two specific sites before its complete digestion within lysosomes. However, unlike its ligand, cycloheximide studies indicated that internalized mIL-3 receptors are recycled to the cell surface. When B6SUtA1 cells were continuously passaged in mIL-3, cell populations allowed to exhaust the mIL-3 in the medium (high density cells) expressed more than ten times (ie, approximately 100,000/cell) the mIL-3 receptor number of those growing exponentially at low cell concentrations (low density cells). Since the high density cells were no larger than the low density cells, the marked increase in mIL-3 receptor number per cell reflects a true up-regulation of receptor expression. A kinetic analysis of this up- regulation revealed that it begins within one hour of mIL-3 exhaustion. Moreover, proliferation assays with these two cell populations, using 3H-thymidine incorporation, suggested that the high density cells were 30-fold more responsive to mIL-3. However, when B6SUtA1 cells were passaged in mGM-CSF, there was no difference in mIL-3 receptor number between high density and low density cells (ie, approximately 100,000/cell). Identical studies carried out with another mIL-3 dependent cell line, 32D C3, demonstrated that this phenomenon was not unique to B6SUtA1 cells.
Human normal CD5+ B lymphocytes can be induced to differentiate to CD5- B lymphocytes with germinal center cell features
Authors: Caligaris-Cappio F , Riva M , Tesio L , Schena M , Gaidano G , Bergui L 
Published: October 2019

Abstract
A significant proportion of cord blood CD5+ B cells express the activation molecules CD23, CD25, and transferrin receptor; react with the cell-cycle-associated monoclonal antibody (MoAb) Ki67; can be induced to enter the S phase of cell cycle by interleukin-2 (IL-2), IL- 4, or low-molecular-weight B-cell growth factor (Imw-BCGF) and, exposed to IL-1 and IL-2, acquire the features (sIgD-, CD5-, CD10+, CD38+) of B blasts proliferating in the germinal centers of secondary follicles. These findings indicate that CD5+ B cells are preactivated and, in the proper microenvironment, may give rise to CD5- B cells.
Growth factors influence the sensitivity of leukemic stem cells to cytosine arabinoside in culture
Authors: Miyauchi J , Kelleher CA , Wang C , Minkin S , McCulloch EA 
Published: October 2019

Abstract
We have proposed that the blasts in acute myeloblastic leukemia (AML) are renewal populations maintained by a small subpopulation of stem cells. The balance between self-renewal and differentiation in blast stem cells may be an important attribute contributing to treatment outcome. Cytosine arabinoside (ara-C) is included in most chemotherapeutic regimens for the treatment of AML. When ara-C survival curves are constructed, the drug appears to be more toxic when an assay is used that detects principally self-renewing divisions, compared with a procedure that depends on terminal divisions. AML blasts usually respond in culture to myelopoietic growth factors; their response often includes a change in self-renewal, differentiation, or both. These features of the model for AML blasts led to the prediction that growth factors would alter ara-C survival curves in a way that depended on the effects of the culture conditions on self-renewal and differentiation. Four AML blast populations were chosen to test this prediction on the basis of our ability to manipulate them by adding or withholding one or more growth factors. Highly significant changes were seen in the ara-C survival curves, depending on the growth factors present in the cultures as was predicted by the observed effects of the factors on renewal and differentiation.
Comparative biochemical and cytogenetic studies of childhood acute lymphoblastic leukemia with the Philadelphia chromosome and other 22q 11 variants
Authors: Dow LW , Tachibana N , Raimondi SC , Lauer SJ , Witte ON , Clark SS 
Published: October 2019

Abstract
We studied the relationship of direct karyotypes, determined at diagnosis and remission, to Abelson-related tyrosine kinase activity and the cytogenetic features of erythroid and myeloid colonies derived from remission marrow of six children with acute lymphoblastic leukemia (ALL). These patients had either the characteristic Philadelphia chromosome (Ph1) [t(9;22)(q34;q11)] or cytogenetically similar variants with a 22q11 breakpoint but no detectable cytogenetic involvement of 9q34. The findings suggested two distinct subtypes of ALL: one defined by t(9;22)(q34;q11) and expression of P185BCR-ABL tyrosine kinase and one with variant karyotypes and no P185BCR-ABL expression. The former comprises cases with Ph1 + marrow cells and Ph1 + erythroid and (or) myeloid colonies in remission marrow and others in which the t(9;22) is undetectable in remission marrow cells. In the latter subgroup, the disease may reflect more extreme mosaicism with a similar stem cell that is cytogenetically undetectable. Variant karyotypes included a del(22)(q11) in one patient and a t(6;22;15;9) (q21;q11;q?22;q21) in another; in both instances, the malignant blast cells lacked P185BCR- ABL expression. Thus ALL with t(9;22)(q34;q11) should be distinguished from ALL with other involvement of the 22q11 breakpoint by molecular studies including protein expression. The diversity of karyotypic findings in cases with involvement of 22q11 suggests at least two mechanisms of leukemogenesis in patients with ALL defined by this breakpoint.
Erythropoietin receptors on murine erythroid colony-forming units: natural history
Authors: Landschulz KT , Noyes AN , Rogers O , Boyer SH 
Published: October 2019

Abstract
Erythropoietin (Epo) response and binding was assessed in purified murine CFU-E and their descendants. Several features emerged. First, Epo on CFU-E is in rapid flux: Half-time for 125I-Epo internalization is approximately four to five minutes. Second, computer-aided Scatchard analyses indicate that greater than 70 high-affinity Epo-receptor sites on anemic animal CFU-E are sometimes already occupied by Epo acquired in vivo. When this is removed, 40% of greater than or equal to 370 sites per CFU-E belong to a high-affinity class (dissociation constant, kd: 73 pmol/L +/- 15 [SE]) and 60% belong to a low-affinity class (kd: 813 pmol/L +/- 246). Third, the few small colonies that develop from CFU-E in the absence of Epo are shown, by serial assay of 59Fe-heme biosynthesis, to stem from contaminating erythroblasts: a result consistent with our finding that, after eight-hour CFU-E culture, most erythroblasts no longer require appreciable Epo for growth. Thus, although the early need for Epo by CFU-E is nearly absolute, this need is not met by the often substantial Epo already on board. The inference is that repeated occupancy of the rapidly turning over Epo receptors is required. Fourth, Epo bound and/or internalized by CFU-E descendants decreases to 40% of zero-time levels after 14 hours in Epo-supplemented culture and disappears after 28 hours. Scatchard analyses indicate that 73 pmol/L kd receptor sites become undetectable at seven to eight hours, whereas 813 pmol/L kd sites are undiminished and only one-third less by 16 hours. This apparent disappearance of high-affinity sites and persistence of low-affinity sites suggests that (a) at least two gene products mediate Epo binding, eg, two different receptor polypeptides or one receptor and one cofactor which modulates affinity; (b) high-affinity sites mediate the growth function of Epo during the first eight hours of culture; and (c) lingering low-affinity receptors may mediate some unrecognized Epo function. Fifth, the efficiency with which 106- and 91-Kd CFU-E membrane polypeptides can be cross-linked to 125I-Epo is two- to threefold higher for cells labeled at high Epo concentrations than at low ones, which suggests that these polypeptides largely reflect low-affinity site reactions.
Nonneoplastic hematopoietic myeloproliferative syndrome induced by dysregulated multi-CSF (IL-3) expression
Authors: Chang JM , Metcalf D , Lang RA , Gonda TJ , Johnson GR 
Published: October 2019

Abstract
Post 5-fluorouracil-treated murine marrow cells were infected with a retroviral vector (MPZen) bearing a multi-potential colony stimulating factor (Multi-CSF) cDNA insert and then transplanted into lethally irradiated syngeneic recipients to study the effects of autocrine production of Multi-CSF in normal hematopoietic cells. Extremely high levels (14,000 U/mL) of Multi-CSF were detected in the sera and in media conditioned by various hematopoietic tissues of the transplanted animals. While spleen, peritoneal, and peripheral blood cellularity increased approximately 10-fold, 10-fold, and 50-fold, respectively, bone marrow cellularity decreased twofold. Progenitor numbers were depressed twofold in the bone marrow but elevated more than 100-fold in the spleen and peritoneum. The majority (80%) of transplanted mice died within 5 weeks of transplantation and showed extensive neutrophilic infiltration of the spleen, lung, liver, and muscle, often with mast cell foci; a phenomenon also seen in the skin and intestine. Neither the infected cells from hematopoietic tissues of the primary mice, nor autonomous mast cell-lines that grew from these cells in liquid culture produced any overt disease when transplanted into normal or sublethally irradiated secondary recipients. In contrast, injection into mice of autonomous FDC-P1 cells transformed by the same retroviral construct led to tumor formation in vivo within 4 weeks. Thus, dysregulated Multi- CSF expression by normal hematopoietic cells produces a fatal but nonneoplastic myeloproliferative syndrome.
Cloning of cDNA coding for connective tissue activating peptide III from a human platelet-derived lambda gt11 expression library
Authors: Wenger RH , Wicki AN , Walz A , Kieffer N , Clemetson KJ 
Published: October 2019

Abstract
We report here the cloning of the cDNA coding for platelet connective tissue-activating peptide-III (CTAP-III) from a lambda gt11 expression library prepared using messenger RNA (mRNA) isolated from human platelets. The open reading frame of the clone coded for a protein with 128 amino acid residues. Since the precursor of CTAP-III, platelet basic protein (PBP is 94 amino acids long, the 5′-translated region of the cDNA codes for a leader sequence 34 amino acids long. This leader sequence, like the sequence of mature CTAP-III, shows significant homology to the sequence of platelet factor 4 (PF4), the only other platelet specific alpha-granule protein cloned until now, from a human erythroleukemic (HEL) cell line-derived cDNA library. These leader sequences are probably critical for targeting such proteins to the alpha-granule. Northern blot hybridization with platelet and megakaryocyte mRNA shows a single species mRNA of approximately 0.8 kb, suggesting that the corresponding cDNA is full length. The cloning of platelet specific CTAP-III provides additional evidence for the platelet specificity of the cDNA library used.
Interaction of fibronectin with cultured human endothelial cells: characterization of the specific receptor
Authors: Conforti G , Zanetti A , Colella S , Abbadini M , Marchisio PC , Pytela R , Giancotti F , Tarone G , Languino LR , Dejana E 
Published: October 2019

Abstract
In this study we provide a characterization of the fibronectin (FN) binding to endothelial cells (EC), and we identify the FN binding site on these cells. 125I-FN binding to EC in suspension was time dependent and reached a plateau at 4 h. Cold FN inhibited this interaction in a concentration-dependent way, but vitronectin, fibrinogen, and IgG were poorly effective. About 80% of the total FN associated to EC at the equilibrium was specifically bound; of this, 60% was reversibly bound, while 20% appeared to be internalized. The FN binding was saturable and an apparent dissociation constant of about 0.23 x 10(-6) mol/L and a maximal number of binding sites of about 9.8 x 10(5) was estimated from binding isotherms. Autoradiography data showed that EC-associated 125I- FN was all in high mol wt form that did not enter the gel. We then characterize the FN receptor (FNR) in EC. An antiserum to the FNR isolated from human placenta inhibited FN binding to EC by 89%, and using the immunoblotting technique, it recognized two bands in the EC detergent extract of mol wt 125/160 Kd. This antiserum also recognized the EC membrane protein complex eluted from the FN affinity column by an arg-gly-asp (RGD) peptide. When this complex was included into liposomes, it poorly bound to FN. However, the binding was strikingly increased by addition of Mn in the buffer and was specific for FN in respect to other substrata. These data define the FN binding site in EC and indicate that it is functionally and structurally related to that isolated from human placenta.
Prolonged defects of interleukin-2 production, responsiveness, and receptor expression in patients with acute lymphoblastic leukemia
Authors: Borzy MS , Ridgway D 
Published: October 2019

Abstract
The proliferative responsiveness to, production of, and the expression of cell-surface receptors for interleukin-2 (IL-2) were examined in 14 children with acute lymphoblastic leukemia (ALL) in remission and receiving maintenance chemotherapy for 6 to 35 months; in 19 children with ALL in remission and off all therapy for 2 to 138 months; and 15 control subjects. Short-term concanavalin A (Con A)-activated, purified T lymphocytes from patients on, as well as patients off, therapy had a significantly decreased proliferative responsiveness to a saturating amount of exogenous, recombinant IL-2 as compared to control subjects (P less than 0.005 and less than 0.05, respectively). Phytohemagglutinin (PHA)-stimulated IL-2 production by peripheral blood mononuclear cells (PBMC) was also substantially decreased in both patient groups with the median values of IL-2 produced being 2.2, 2.1, and 8.1 U/mL in the on therapy, off therapy, and control groups, respectively. In addition, PHA-induced expression of cell-surface receptors for IL-2 on PBMC was significantly decreased in both patient groups as compared to control subjects (P less than 0.01). Lymphocyte proliferation to mitogens (PHA, Con A, and pokeweed mitogen) was similar in all three groups studied. These results demonstrate that substantial quantitative and qualitative abnormalities of the IL-2-T lymphocyte system are present in the majority of treated patients with ALL, not only during maintenance therapy, but also for a prolonged period after the cessation of all chemotherapy. These long-lasting defects of the IL-2 system are most likely a late effect of chemotherapy and may result in increased complications in some long- term survivors of ALL.
Extranodal noncutaneous lymphoid hyperplasias represent a continuous spectrum of B-cell neoplasia: demonstration by molecular genetic analysis
Authors: Knowles DM , Athan E , Ubriaco A , McNally L , Inghirami G , Wieczorek R , Finfer M , Jakobiec FA 
Published: October 2019

Abstract
We investigated 16 lymphoid proliferations occurring in the ocular adnexa, salivary glands, breast, and thyroid gland and satisfying the histopathologic and immunophenotypic criteria of benign lymphoid hyperplasia for the presence of clonal rearrangements of the antigen receptor, c-myc, bcl-1, and bcl-2 genes and Epstein-Barr virus (EBV) DNA sequences. Each of these 16 extranodal, noncutaneous lymphoid neoplasms exhibited clonal immunoglobulin heavy and/or light chain and lacked T-cell receptor (TCR) beta-chain gene rearrangements. The patterns of immunoglobulin gene rearrangements included solitary and multiple barely perceptible to faint bands, solitary clear and definite bands, and solitary high-intensity bands superimposed on a background of multiple less-intense bands. Three ocular adnexal lymphoid neoplasms exhibited bcl-1 or bcl-2 gene rearrangements. None of the 16 lymphoid neoplasms contained EBV DNA sequences. Two patients developed a histopathologically confirmed malignant lymphoma in an extranodal site. None of the remaining 14 patients developed additional lymphoid neoplasms during a mean follow-up period of 30 months, despite conservative therapy. These results demonstrate that extranodal, noncutaneous lymphoid neoplasms meeting the histopathologic and immunophenotypic criteria for benign lymphoid hyperplasia frequently contain occult monoclonal and oligoclonal B-cell populations representing a continuous and progressive spectrum of B-cell neoplasia up to and including malignant lymphoma.
A beta-thalassemia gene caused by a 290-base pair deletion: analysis by direct sequencing of enzymatically amplified DNA
Authors: Spiegelberg R , Aulehla-Scholz C , Erlich H , Horst J 
Published: October 2019

Abstract
The base composition around a recently detected deletion in the human beta-globin gene was determined by direct DNA sequencing of an enzymatically amplified DNA segment. The deletion removes 290 base pairs (bp), including the entire exon 1 and the mRNA cap site. In the vicinity of the deletion endpoints, the normal beta-globin gene contains direct and inverted repeats which may have taken part in generation of this deletion.
A C----T substitution at nt--101 in a conserved DNA sequence of the promotor region of the beta-globin gene is associated with “silent” beta-thalassemia
Authors: Gonzalez-Redondo JM , Stoming TA , Kutlar A , Kutlar F , Lanclos KD , Howard EF , Fei YJ , Aksoy M , Altay C , Gurgey A 
Published: October 2019

Abstract
Sequence analyses and dot-blot analyses with synthetic oligonucleotide probes have identified eight individuals in three Turkish families and one Bulgarian family with one chromosome having a C----T mutation at nucleotide position--101 relative to the Cap site of the beta-globin gene. This nucleotide is part of one of the conserved blocks of nucleotides within the promoter region; in vitro expression analyses with the chloramphenicol acetyltransferase system showed that this substitution will decrease the effectiveness of transcription. Five subjects had a thalassemia intermedia due to the additional presence of a known classical high hemoglobin (Hb) A2 beta-thalassemia mutation on the second chromosome; their hematologic condition was relatively mild. The three persons with a heterozygosity for the--101 C----T mutation had normal hematologic data without microcytosis but with high-normal levels of Hb A2 and a mild imbalance in chain synthesis. The newly discovered mutation is considered one of the silent types of beta- thalassemia. It is relatively rare because it was absent among several hundred normal and beta-thalassemia chromosomes.