Identification of PDGF Receptors on Human Megakaryocytes and Megakaryocytic Cell Lines

M Yang; L M Khachigian; C Hicks; C N Chesterman; B H Chong

doi:10.1055/s-0038-1657648

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1997; 78(02): 892-896
DOI: 10.1055/s-0038-1657648

Rapid Communication

Schattauer GmbH Stuttgart

Identification of PDGF Receptors on Human Megakaryocytes and Megakaryocytic Cell Lines

M Yang

The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia

,

L M Khachigian

The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia

,

C Hicks

The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia

,

C N Chesterman

The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia

,

B H Chong

The Centre for Thrombosis and Vascular Research, School of Pathology, The University of New South Wales and Department of Haematology, The Prince of Wales Hospital, Sydney, Australia

› Author Affiliations

Abstract

PDF Download

Summary

Platelet-derived growth factor (PDGF) is a potent chemotactic and mitogenic factor implicated to play important roles in a variety of normal and pathophysiologic settings. We investigated PDGF receptor expression on human megakaryocytes and several megakaryocytic cell lines (CHRF, DAMI, Meg-01, M-07e) using enzyme-linked immunosorbent assay (ELISA), flow cytometry and immunocytochemical staining. Both PDGF receptor subtypes were identified on CHRF, DAMI, and Meg-01 cells by ELISA; PDGF beta-receptor levels exceeded alpha-receptor levels. Flow cytometry revealed that beta-receptor levels on CHRF and DAMI cells exceeded those on Meg-01 cells, and that M-07e expressed neither receptor. Immunocytochemical staining confirmed these findings and determined that bone marrow megakaryocytes also expressed PDGF receptors. Exposure of megakaryocytes to PDGF-BB dramatically induced the expression of the immediate-early gene, c-fos, within 30 min. Moreover, PDGF-BB significantly stimulated CHRF proliferation and colony formation. The present study demonstrates the presence of functional PDGF receptors on human megakaryocytes and their ability to mediate a mitogenic response.

PDF (382 kb)

References

References
1 Claesson-Welsh L, Hammacher A, Westermark B, Heldin CH, Nister M. Identification and structural analysis of the A type receptor for platelet- derived growth factor: similarities with the B type receptor. J Biol Chem 1989; 264: 1742-1747
2 Gronwald RGK, Grant FJ, Haldeman BA, O’Hara PJ, Hagen FS, Ross R, Bowen-Pope DF, Murry MJ. Cloning and expression of a cDNA coding for the human platelet-derived growth factor receptor: evidence for more than one receptor class. Proc Natl Acad Sci USA 1988; 85: 3435-3439
3 Matsui T, Heidaran M, Miki T, Popesal N, La RochelleW, Kraus M, Pierce G, Aaronson S. Isolation of a novel receptor cDNA establishes the existence of two PDGF receptor genes. Science 1989; 243: 800-804
4 Gladwin AM, Carrier MJ, Beesley JE, Lelchuk R, Hancock V, Martin JF. Identification of mRNA for PDGF B-chain in human megakaryocytes isolated using a novel immunomagnetic separation method. Br J Haematol 1990; 76: 333-339
5 Cheng S, Rifas L, Shen V, Tong B, Pierce G, Deuel T, Peck W. J774A.1 macrophage cell line produces PDGF-like and non-PDGF-like growth factors for bone cells. J Bone Miner Res 1987; 02: 467-474
6 Malden L, Chait A, Raines E, Ross R. The influence of oxidatively modified low density lipoproteins on expression of platelet-derived growth factor by human monocyte derived macrophages. J Biol Chem 1991; 266: 13901-13907
7 Ross R, Masudea J, Raines E, Gown A, Katsuda S, Sasahara M, Malden L, Masuko H, Sato H. Localization of PDGF-B protein in macrophages in all phases of atherogenesis. Science 1990; 248: 1009-1012
8 Sariban E, Kufe D. Expression of the platelet-derived growth factor 1 and 2 genes in human myeloid cell lines and monocytes. Cancer Res 1988; 48: 4498-4502
9 Deuel T, Kawahara R, Mustoe T, Pierce A. Growth factors and wound healing: platelet-derived growth factor as a model cytokine. Annu Rev Med 1991; 42: 567-574
10 Tsai LH, White L, Raines E, Ross R, Smith RG, Cushley W, Ozanne B. Expression of platelet-derived growth factor and its receptors by two Pre-B acute lymphocytic leukemia cell lines. Blood 1994; 85: 51-55
11 Yang M, Chesterman CN, Chong BH. Effect of PDGF on murine granu- locyte-macrophage colony formation in vitro. Exp Haematol 1994; 22: 738 Abstract
12 Yang M, Chesterman CN, Chong BH. Recombinant PDGF enhances megakaryocytopoiesis in vitro. Br J Haematol 1995; 91: 285-289
13 Dainiak N, Davis G, Kalmanti M, Lawler J, Kulkami V. Platelet-derived growth factor promotes proliferation of erythropoietic progenitor cells in vitro. J Clin Invest 1983; 71: 1206-1214
14 Soriano P. Abnormal kidney development and hematological disorders in PDGF beta-receptor mutant mice. Genes & Develop 1994; 8: 1888-1896
15 Leveen P, Pekny M, Gebre-Medhin S, Swolin B, Larsson E, Betsholtz C. Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities. Genes & Develop 1994; 08: 1875-1887
16 Goustin AS, Galanopoulos T, Kalyanaraman VS, Pantazis P. Co-expression of the genes for platelet-derived growth factor and its receptor in human T-cell lines infected with HTLV-I. Growth Factors 1990; 02: 189-195
17 Pantazis P, Goustin AS, Nixon J. Platelet-derived growth factor and its receptor in blood cell differentiation and neiplasia. Eur J Haematol 1990; 45: 127-138
18 Vassbotn FS, Havnen OK, Heldin C, Holmson H. Negative feedback regulation of human platelets via autocrine activation of the platelet-derived growth factor alpha-receptor. J Biol Chem 1994; 269: 13874-13879
19 Ogura M, Morishima Y, Ohno R, Kato Y, Hirabayashi N, Nagura H, Saito H. Establishment of a novel human megakaryoblastic leukemia cell line, MEG-01, with positive Philadelphia chromosome. Blood 1985; 66: 1384-1392
20 Hicks C, Sleigh MJ, Chesterman CN. Quantitation of platelet-derived growth factor receptors on cells from human arterial segments: J Immunol Methods. 1994; 170: 83-92
21 Wu ZH, Chesterman CN, Chong BH. Identification of IgG Fc receptor type II on human megakaryoblastic cell lines (MEG-01 and UT-7). Br J Haematol 1993; 84: 204-211
22 Siefert RA, Hart CE, Phillips PE, Forstrom JW, Ross R, Murray MJ, Bowen-Pope DF. Two different subunits associate to create isoform- specific platelet-derived growth factor receptors. J Biol Chem 1989; 264: 8771-8778
23 Heldin CH, Westermark B. Platelet-derived growth factors: a family of isoforms that bind to two distinct receptors. Br Med Bull 1989; 45: 453-464
24 Bar RS, Boes M, Booth BA, Dake BL, Henley S, Hart MN. The effects of platelet-derived growth factor in cultured micro vessel endothelial cells. Endocrinology 1989; 124: 1841-1848
25 Smits A, Hermansson M, Nister M, Kamushina I, Heldin CH, Westermark B, Funa K. Rat brain capillary endothelial cells express functional PDGF-B type receptor. Growth Factors 1989; 02: 01-08
26 Smits A, Kato M, Westermark B, Nister M, Heldin CH, Funa K. Neurotro- phil activity of platelet-derived growth factor: Rat neuronal cells possess functional PDGF beta-type receptors and respond to PDGF. Proc Natl Acad Sci USA 1991; 88: 8159-8163