Showing papers by "William J. Murphy published in 1999"

Vascular Endothelial Growth Factor and Basic Fibroblast Growth Factor Induce Expression of CXCR4 on Human Endothelial Cells: In Vivo Neovascularization Induced by Stromal-Derived Factor-1α

Abstract: The contribution of chemokines toward angiogenesis is currently a focus of intensive investigation. Certain members of the CXC chemokine family can induce bovine capillary endothelial cell migration in vitro and corneal angiogenesis in vivo, and apparently act via binding to their receptors CXCR1 and CXCR2. We used an RNAse protection assay that permitted the simultaneous detection of mRNA for various CXC chemokine receptors in resting human umbilical vein endothelial cells (HUVECs) and detected low levels of only CXCR4 mRNA. Stimulation of HUVECs with vascular endothelial growth factor (VEGF) or basic fibroblast growth factor (bFGF) up-regulated levels of only CXCR4 mRNA. CXCR4 specifically binds the chemokine stromal-derived factor-1α (SDF-1α). Competitive binding studies using 125I-labeled SDF-1α with Scatchard analysis indicated that VEGF or bFGF induced an average number of approximately 16,600 CXCR4 molecules per endothelial cell, with a Kd = 1.23 × 10−9 mol/L. These receptors were functional as HUVECs and human aorta endothelial cells (HAECs) migrated toward SDF-1α. Although SDF-1α-induced chemotaxis was inhibited by the addition of a neutralizing monoclonal CXCR4 antibody, endothelial chemotaxis toward VEGF was not altered; therefore, the angiogenic effect of VEGF is independent of SDF-1α. Furthermore, subcutaneous SDF-1α injections into mice induced formation of local small blood vessels that was accompanied by leukocytic infiltrates. To test whether these effects were dependent on circulating leukocytes, we successfully obtained SDF-1α-induced neovascularization from cross sections of leukocyte-free rat aorta. Taken together, our data indicate that SDF-1α acts as a potent chemoattractant for endothelial cells of different origins bearing CXCR4 and is a participant in angiogenesis that is regulated at the receptor level by VEGF and bFGF.

The Promise of Comparative Genomics in Mammals

Abstract: Dense genetic maps of human, mouse, and rat genomes that are based on coding genes and on microsatellite and single-nucleotide polymorphism markers have been complemented by precise gene homolog alignment with moderate-resolution maps of livestock, companion animals, and additional mammal species. Comparative genetic assessment expands the utility of these maps in gene discovery, in functional genomics, and in tracking the evolutionary forces that sculpted the genome organization of modern mammalian species.

Extensive conservation of sex chromosome organization between cat and human revealed by parallel radiation hybrid mapping.

Abstract: A radiation hybrid (RH)-derived physical map of 25 markers on the feline X chromosome (including 19 Type I coding loci and 6 Type II microsatellite markers) was compared to homologous marker order on the human and mouse X chromosome maps. Complete conservation of synteny and marker order was observed between feline and human X chromosomes, whereas the same markers identified a minimum of seven rearranged syntenic segments between mouse and cat/human X chromosome marker order. Within the blocks, the feline, human, and mouse marker order was strongly conserved. Similarly, Y chromosome locus order was remarkably conserved between cat and human Y chromosomes, with only one marker (SMCY) position rearranged between the species. Tight linkage and a conserved gene order for a segment encoding three genes, DFFRY-DBY-UTY in human, mouse, and cat Y chromosomes, coupled with demonstrated deletion effects of these genes on reproductive impairment in both human and mouse, implicates the region as critical for Y-mediated sperm production.

Tyrphostin AG-490 inhibits cytokine-mediated JAK3/STAT5a/b signal transduction and cellular proliferation of antigen-activated human T cells.

Abstract: Janus kinase 3 (JAK3) is a cytoplasmic tyrosine kinase required for T cell development and activated by cytokines that utilize the interleu- kin-2 (IL-2) receptor common gamma chain (gc). Genetic inactivation of JAK3 is manifested as se- vere combined immunodeficiency disease (SCID) in humans and mice. These findings have suggested that JAK3 represents a pharmacological target to control certain lymphoid-derived diseases. Here we provide novel evidence that AG-490 potently inhib- its the autokinase activity of JAK3 and tyrosine phosphorylation and DNA binding of signal trans- ducer and activator of transcription 5a and 5b (STAT5a/b). Similar inhibitory effects were ob- served with other cytokines that use gc. AG-490 also inhibited IL-2-mediated proliferative growth in human T cells with an IC50 5 25 mM that was partially recoverable. Moreover, we demonstrate that this inhibitor prevented tetanus toxoid antigen- specific T cell proliferation and expansion but failed to block activation of Zap70 or p56Lck after anti-CD3 stimulation of human T cells. Taken together, these findings suggest that AG-490 inhib- its the JAK3-mediated Type II signaling pathway but not the T cell receptor-derived Type I pathway and possesses therapeutic potential for T cell- derived pathologies such as graft-versus-host dis- ease, allergy, and autoimmune disorders. J. Leu- koc. Biol. 65: 891-899; 1999.

Ly-49 receptor expression and functional analysis in multiple mouse strains

Abstract: We present data on the strain distribution and functional characteristics of the Ly-49 receptors A, C/I, D, and G2 on DX5+ natural killer (NK) cells. We have examined tyrosine phosphorylation of the Ly-49 molecules, regulation of NK cytotoxic functions, and in vivo marrow rejection capability. The flow cytometry results demonstrate a diverse and complex pattern of expression of the Ly-49 receptors in the 11 strains examined. The vast majority of NK cells express Ly-49s, although some NK1.1+ CD3+ cells also express these receptors. The results of our functional analysis indicate that H-2Dd was able to inhibit the function of Ly-49G2+ NK cells, not only in B6 mice, but also by NK cells derived from several haplotypes. The examination of Ly-49 receptor tyrosine phosphorylation, which is a biochemical measure of inhibitory function, was consistently observed in the 11 mouse strains examined. In contrast, analysis of Ly-49D function suggests its expression appears to be more restricted and that H-2Dd is an activating ligand for this receptor. In addition, the in vivo examination of both inhibitory (Ly-49G2) and activating (Ly-49D) receptors demonstrated regulatory roles of these class I binding receptors in marrow transplantation.

Genome maps 10. Comparative genomics. Mammalian radiations. Wall chart.

Abstract: ![Figure][1] Traces of evolutionary history appear in functional morphology and DNA sequences of living and extinct species These remnants of the past can lead to insights into the relationships among extant groups of animals, the forces driving evolution, and the utility of animal models for studying human disease We present below one evolutionary interpretation of the still-disputed hierarchy of surviving placental mammalian orders (excluding monotremes and marsupials), a synthesis of accumulated molecular and morphological inferences The time scale is derived largely from molecular data; indicated fossil remains are much younger, raising controversies around the precise age of mammal ancestors The tips of the phylogenetic tree depict genomes for 21 representative species from 11 orders, which are labeled on the vertical lines These examples show genome-wide homology alignments assessed by comparative gene mapping or direct visualization of chromosome painting, in which fluorescently labeled, individual chromosomes are hybridized to chromosomes from distantly related species Species were selected to maximize the number of orders represented and to illustrate patterns of genome conservation The 24 human chromosomes are distinguished by 24 colors, and regions of human chromosome homology in other species are indicated by color and human chromosome number (for example, cat chromosome A1 contains gene stretches homologous to human chromosomes 5 and 13) Selected gene homologs in each species represent a subset of extensive comparative gene mapping data; thus genes listed may not reflect the extreme borders of the conserved segments Gene orders have been determined in human, cat, pig, sheep, rat, mouse, and zebrafish; in other species the genes are listed in the order in which they appear in humans The zebrafish map, including 25 linkage groups not yet assigned to the 25 chromosomes, is presented to illustrate the array of gene segments conserved for 450 million years Further information and discussion of dissenting views of mammalian evolution can be found in this issue of Science and the associated web site at ![Figure][1] CREDITS Science Coordinators: Barbara R Jasny and Pamela J Hines Authors: Phylogenetic tree Stephen J O'Brien, National Cancer Institute, Frederick, MD, USA; John F Eisenberg, University of Florida, Gainesville, FL, USA; Michael Miyamoto, University of Florida, Gainesville, FL, USA; S Blair Hedges, Pennsylvania State University, University Park, PA, USA; Sudhir Kumar, Arizona State University, Tempe, AZ, USA; Don E Wilson, Smithsonian Institution, Washington, DC, USA Genomic maps Stephen J O'Brien, Marilyn Menotti-Raymond, William J Murphy, William G Nash, Leslie A Lyons, Joan C Menninger, Roscoe Stanyon, Johannes Wienberg, Neal G Copeland, Nancy A Jenkins, National Cancer Institute, Frederick, MD, USA; Joel Gellin, Martine Yerle, Institut National de la Recherche Agronomique, Castanet-Tolosan, France; Leif Andersson, Swedish University of Agricultural Sciences, Uppsala, Sweden; James Womack, Texas A & M University, College Station, TX, USA; Thomas Broad, AgResearch, Invermay, Mosgiel, New Zealand; John Postlethwait, University of Oregon, Eugene, OR, USA; Oleg Serov, Institute of Cytology and Genetics, Siberian Branch of the Academy of Sciences of Russia, Novosibirsk, Russia; Ernie Bailey, University of Kentucky, Lexington, KY, USA; Michael R James, Wellcome Trust Centre for Human Genetics, Headington, UK; Takeshi K Watanabe, Otsuka GEN Research Institute, Tokushima, Japan; Matthew J Wakefield, Jennifer Marshall Graves, La Trobe University, Melbourne, Australia Design and Art Direction: C Faber Smith Graphics: Nathalie Cary Illustrations: Katharine Sutliff Production Assistance: Holly Bishop, Leslie Blizard, Inja Lin, Darcel Pugh Proofreading: Harry Jach [1]: pending:yes

Use of neuroendocrine hormones to promote reconstitution after bone marrow transplantation.

Abstract: A survey of the previous literature and the data shown here indicate that neuroendocrine hormones such as growth hormone and prolactin may be of potential clinical use after bone marrow transplantation (BMT) to promote hematopoietic and immune recovery. The amounts of hormones used in our model do not promote weight gain suggesting that their lymphohematopoietic actions were independent of their anabolic effects. While the hormones may not produce the same extent of immune/hematopoietic effects when compared to conventional hematopoietic and immune stimulating cytokines (i.e. IL-2 or G-CSF), their pleiotropic effects and limited toxicity after systemic administration makes them attractive to test in the post-BMT setting. However, more work needs to be performed to understand the mechanism(s) of their action, particularly with regard to T-cell function and development.

The promise of comparative genomics in mammals : Genome

Abstract: Dense genetic maps of human, mouse, and rat genomes that are based on coding genes and on microsatellite and single-nucleotide polymorphism markers have been complemented by precise gene homolog alignment with moderate-resolution maps of livestock, companion animals, and additional mammal species. Comparative genetic assessment expands the utility of these maps in gene discovery, in functional genomics, and in tracking the evolutionary forces that sculpted the genome organization of modern mammalian species.