Download PDF
Thromb Haemost 1993; 70(02): 273-280
DOI: 10.1055/s-0038-1649565
Original Articles
Coagulation
Schattauer GmbH Stuttgart

Characterization of Monocyte-Associated Factor V

Janos Kappelmayer
The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, USA
,
Satya P Kunapuli
The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, USA
,
Edward G Wyshock
The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, USA
,
Robert W Colman
The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, USA
› Author Affiliations
Further Information

Publication History

Received 20 July 1992

Accepted after revision 02 February 1993

Publication Date:
04 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

We demonstrate that in addition to possessing binding sites for intact factor V (FV), unstimulated peripheral blood monocytes also express activated factor V (FVa) on their surfaces. FVa was identified on the monocyte surface by monoclonal antibody B38 recognizing FVa light chain and by human oligoclonal antibodies H1 (to FVa light chain) and H2 (to FVa heavy chain) using immunofluorescence microscopy and flow cytometry. On Western blots, partially cleaved FV could be identified as a 220 kDa band in lysates of monocytes. In addition to surface expression of FVa, monocytes also contain intracellular FV as detected only after permeabilization by Triton X-100 by monoclonal antibody B10 directed specifically to the Cl domain not present in FVa. We sought to determine whether the presence of FV in peripheral blood monocytes is a result of de novo synthesis.

Using in situ hybridization, no FV mRNA could be detected in monocytes, while in parallel control studies, factor V mRNA was detectable in Hep G2 cells and CD18 mRNA in monocytes. In addition, using reverse transcriptase and the polymerase chain reaction, no FV mRNA was detected in mononuclear cells or in U937 cells, but mRNA for factor V was present in Hep G2 cells using the same techniques. These data suggest that FV is present in human monocytes, presumably acquired by binding of plasma FV, and that the presence of this critical coagulation factor is not due to de novo synthesis.

 

  • References

  • 1 Dahlbäck B. A new model for coagulation factor V suggesting a unique mechanism of activation. Scand J Clin Lab Invest 1988; 48 (Suppl. 191) 47-61
    CrossrefPubMedGoogle Scholar
  • 2 Miletich JP, Jackson CM, Majerus PW. Properties of the factor Xa binding site on human platelets. J Biol Chem 1978; 253: 6908-6916
    PubMedGoogle Scholar
  • 3 Tracy PB. Regulation of thrombin generation at cell surfaces. Semin Thromb Hemostas 1988; 14: 227-233
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Gewirtz AM, Keefer M, Doshi K, Annamalai A, Chiu HC, Colman RW. Biology of human megakaryocyte factor V. Blood 1986; 67: 1639-1648
    PubMedGoogle Scholar
  • 5 Chiu HC, Schick PK, Colman RW. Biosynthesis of factor V in isolated guinea pig megakaryocytes. J Clin Invest 1985; 75: 339-346
    CrossrefPubMedGoogle Scholar
  • 6 Mazzorana M, Cornillou B, Baffet G, Hubert N, Eloy R, Guguen-Guilluozo C. Biosynthesis of factor V by normal adult rat hepatocytes. Thromb Res 1989; 54: 655-675
    CrossrefPubMedGoogle Scholar
  • 7 Wilson DB, Salem HH, Mruk JS, Maruyama I, Majerus PW. Biosynthesis of coagulation factor V by a human hepatocellular carcinoma cell line. J Clin Invest 1984; 73: 654-658
    CrossrefPubMedGoogle Scholar
  • 8 Tracy PB, Rohrbach MS, Mann KG. Functional prothrombinase complex assembly on isolated monocytes and lymphocytes. J Biol Chem 1983; 258: 7264-7267
    PubMedGoogle Scholar
  • 9 Tracy PB, Eide LL, Mann KG. Human prothrombinase complex assembly and function on isolated peripheral blood cell populations. J Biol Chem 1985; 260: 2119-2124
    PubMedGoogle Scholar
  • 10 Recalde HR. A simple method of obtaining monocytes in suspension. J Immunol Methods 1984; 69: 71-77
    CrossrefPubMedGoogle Scholar
  • 11 Annamalai AE, Stewart GJ, Hansel B, Memoli M, Chiu HC, Manuel DW, Doshi K, Colman RW. Expression of factor V on human umbilical vein endothelial cells is modulated by cell injury. Arteriosclerosis 1986; 6: 196-202
    CrossrefPubMedGoogle Scholar
  • 12 Annamalai AE, Rao AK, Chiu HC, Wang D, Dutta-Roy AK, Walsh PN, Colman RW. Epitope mapping of functional domains of human factor Va with human and murine monoclonal antibodies. Evidence for the interaction of heavy chain with factor Xa and calcium. Blood 1987; 70: 139-146
    PubMedGoogle Scholar
  • 13 Kane WH, Davie E. Cloning of cDNA coding for human factor V, a blood coagulation factor homologous to factor VIII and ceruloplamin. Proc Natl Acad Sci USA 1986; 83: 6800-6804
    CrossrefPubMedGoogle Scholar
  • 14 Feinberg AP, Vogelstein B. A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1984; 137: 266-267
    CrossrefPubMedGoogle Scholar
  • 15 Rappolee DA, Mark D, Banda MJ, Werb Z. Wound macrophages express TGF-alpha and other growth factors in vivo: Analysis by mRNA phenotyping. Science 1988; 241: 708-712
    CrossrefPubMedGoogle Scholar
  • 16 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Lab, Cold Spring Harbor, NY: 1989
    Google Scholar
  • 17 Fuqua SA, Fitzgerald SD, McGuire WL. A simple polymerase chain reaction method for detection and cloning of low-abundance transcripts. Biotechniques 1990; 9: 206-211
    PubMedGoogle Scholar
  • 18 Muszbek L, Adany R, Szegedi G, Polgar F, Kavai M. Factor XIII of blood coagulation in human monocytes. Thromb Res 1985; 37: 401-410
    CrossrefPubMedGoogle Scholar
  • 19 Niemetz F, Morrison DC. Lipid A as the biologically active moiety in bacterial endotoxin (LPS) initiated generation of procoagulant activity in peripheral blood leukocytes. Blood 1977; 49: 947-956
    PubMedGoogle Scholar
  • 20 Carlesen H, Prydz H. Enhancement of procoagulant activity in stimulated mononuclear blood cells and monocytes by cyclosporine. Transplantation 1981; 43: 543-548
    PubMedGoogle Scholar
  • 21 Osterud B, Rapaport SI. Activation of factor IX by the reaction product of tissue factor and factor VII: Additional pathway for initiating blood coagulation. Proc Natl Acad Sci USA 1977; 74: 5260-5264
    CrossrefPubMedGoogle Scholar
  • 22 Owren P. Parahemophilia, hemorrhagic diathesis due to absence of previously unknown clotting factor. Lancet 1947; 1: 6-8
    PubMedGoogle Scholar
  • 23 Tracy PB, Eide LL, Bowie EJ, Mann KG. Radioimmunoassay of factor V in human plasma and platelets. Blood 1982; 60: 59-63
    PubMedGoogle Scholar
  • 24 Cerveny TJ, Fass DN, Mann KG. Synthesis of coagulation factor V by cultured aortic endothelium. Blood 1984; 63: 1467-1474
    PubMedGoogle Scholar
  • 25 Giddings FC, Jarvis AL, Hogg S. Factor V in human vascular endothelium and in endothelial cells in culture. Thromb Res 1986; 44: 829-835
    CrossrefPubMedGoogle Scholar
  • 26 Schwartz BS, Levy GA, Fair DS, Edgington TS. Murine lymphoid procoagulant activity induced by bacterial lipopolysaccharide and immune complexes is a monocyte prothrombinase. J Exp Med 1982; 155: 1464-1479
    CrossrefPubMedGoogle Scholar
  • 27 Hogg N. Human monocytes are associated with the formation of fibrin. J Exp Med 1983; 157: 473-485
    CrossrefPubMedGoogle Scholar
  • 28 Hogg N. Human monocytes have prothrombin cleaving activity. Clin Exp Immunol 1985; 53: 725-730
    PubMedGoogle Scholar
  • 29 Altieri DC, Edgington TS. Sequential receptor cascade for coagulation proteins on monocytes. J Biol Chem 1989; 264: 2969-2972
    PubMedGoogle Scholar
  • 30 Tracy PB, Mann KG. Prothrombinase complex assembly on the platelet surface is mediated through the 74,000 dalton component of factor Va. Proc Natl Acad Sci USA 1983; 80: 2380-2384
    CrossrefPubMedGoogle Scholar
  • 31 Kane WH, Ichinose A, Hagen FS, Davie EW. Cloning of cDNAs coding for the heavy chain region and connecting region of human factor V, a blood coagulation factor with four types of internal repeats. Biochemistry 1987; 26: 6508-6514
    CrossrefPubMedGoogle Scholar
  • 32 Robinson RA, Worfolk L, Tracy PB. Endotoxin enhances the expression of monocyte prothrombinase activity. Blood 1992; 79: 406-416
    PubMedGoogle Scholar
  • 33 Jenny JR, Pittman DD, Toole JJ, Kriz RW, Aldape RA, Hewick RM, Kaufman RJ, Mann KG. Complete cDNA and derived amino acid sequence of human factor V. Proc Natl Acad Sci USA 1987; 84: 4846-4850
    CrossrefPubMedGoogle Scholar
  • 34 Chelly J, Concordet JP, Kaplan JC, Kahn A. Illegitimate transcription: Transcription of any gene in any cell type. Proc Natl Acad Sci USA 1989; 86: 2617-2621
    CrossrefPubMedGoogle Scholar
  • 35 Brenner CA, Tam AW, Nelson PA, Engelman EG, Suzuki N, Frey KE, Larrick JW. Message amplification phenotyping (MAPPing), a technique to simultaneously measure multiple mRNAs from small number of cells. BioTechniques 1989; 7: 1096-1103
    PubMedGoogle Scholar
  • 36 Gregory SA, Morrissey JH, Edgington TS. Regulation of the tissue factor gene expression in the monocyte procoagulant response to endotoxin. Mol Cell Biol 1989; 9: 2752-2755
    CrossrefPubMedGoogle Scholar
  • 37 Kappelmayer J, Adany R. Factors of the extrinsic pathway of blood coagulation in fetal placental macrophages. Am J Reprod Immun 1989; 21: 37-40
    CrossrefPubMedGoogle Scholar
  • 38 Adany R, Kappelmayer J, Berenyi E, Szegedi A, Fabian E, Muszbek L. Factors of the extrinsic pathway of blood coagulation in tumour associated macrophages. Thromb Haemostas 1989; 62: 850-855
    PubMedGoogle Scholar
  • 39 Osterud B, Bogwald J, Lindahl U, Seljelid R. Production of blood coagulation factor V and tissue thromboplastin by macrophages in vitro. FEBS Lett 1981; 127: 154-156
    CrossrefPubMedGoogle Scholar
  • 40 Osterud B, Lindahl U, Seljelid R. Macrophages produce blood coagulation factors. FEBS Lett 1980; 120: 41-43
    CrossrefPubMedGoogle Scholar
  • 41 Rothberger H, McGee MP. Generation of coagulation factor V activity by cultured rabbit alveolar macrophages. J Exp Med 1984; 160: 1880-1890
    CrossrefPubMedGoogle Scholar