Thromb Haemost 2005; 93(01): 160-164
DOI: 10.1160/TH03-11-0703
New Technologies and Diagnostic Tools
Schattauer GmbH

Generation of a polyclonal rabbit anti-mouse tissue factor antibody by nucleic acid immunisation

Andrew D. Mumford
1   Haemostasis and Thrombosis, MRC Clinical Sciences Centre, London, United Kingdom
,
Daxin Chen
2   Department of Immunology, The Faculty of Medicine, Imperial College, Hammersmith Hospital Campus, London, United Kingdom
,
Anthony Dorling
2   Department of Immunology, The Faculty of Medicine, Imperial College, Hammersmith Hospital Campus, London, United Kingdom
,
Geoffrey Kemball-Cook
1   Haemostasis and Thrombosis, MRC Clinical Sciences Centre, London, United Kingdom
,
John H. McVey
1   Haemostasis and Thrombosis, MRC Clinical Sciences Centre, London, United Kingdom
› Author Affiliations
Financial support: The Medical Research Council of the United Kingdom supported this work. ADM was supported by the British Heart Foundation
Further Information

Correspondence to:

Dr John McVey
Haemostasis and Thrombosis
MRC Clinical Sciences Centre
The Faculty of Medicine, Imperial College
Hammersmith Hospital Campus
Du Cane Road
London, W12 0NN
United Kingdom
Phone: 020 8383 8253   
Fax: 0870 131 3540   

Publication History

Received 19 November 2003

Accepted after revision 12 October 2004

Publication Date:
14 December 2017 (online)

 

Summary

Tissue factor (TF) the cellular receptor and cofactor for factor VII, initiates coagulation and has also been implicated in several coagulation-independent functions, including inflammation, angiogenesis and tumour metastasis. Investigations of TF expression in mouse models of these processes has been limited by the availability of antibodies that specifically recognise mouse TF.We have generated a rabbit polyclonal antibody to mTF by DNA immunisation. This has yielded an antiserum that recognises native mTF in immunohistochemical and flow cytometric analyses.Furthermore,the antiserum is inhibitory in coagulation assays.This antiserum will be a valuable investigative tool in the analysis of mTF expression.


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  • References

  • 1 Kemball-Cook G, Johnson DJD, Takamiya O. et al Coagulation factor VII Gln100-->Arg – Amino acid substitution at the epidermal growth factor 2-protease domain interface results in severely reduced tissue factor binding and procoagulant function. J Biol Chem 1998; 273: 8516-21.
  • 2 Dackiw APB, McGilvray ID, Woodside M. et al Prevention of endotoxin-induced mortality by antitissue factor immunization. Arch Surg 1996; 131: 1273-8.
  • 3 Chen D, Giannopoulos K, Shiels PG. et al Inhibition of intravascular thrombosis in murine endotoxemia by targeted expression of hirudin and tissue factor pathway inhibitor analogs to activated endothelium. Blood 2004; 104: 1344-9.
  • 4 Wolff JA, Malone RW, Williams P. et al Direct gene transfer into mouse muscle in vivo. Science 1990; 247: 1465-8.
  • 5 Wilcox JN, Smith KM, Schwartz SM. et al Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A 1989; 86: 2839-43.
  • 6 Archipoff G, Beretz A, Freyssinet JM. et al Heterogeneous regulation of constitutive thrombomodulin or inducible tissue-factor activities on the surface of human saphenous-vein endothelial cells in culture following stimulation by interleukin-1, tumour necrosis factor, thrombin or phorbol ester. Biochem J 1991; 273: 679-84.
  • 7 Galdal KS, Lyberg T, Evensen SA. et al Thrombin induces thromboplastin synthesis in cultured vascular endothelial cells. Thromb Haemost 1985; 54: 373-6.
  • 8 Brox JH, Osterud B, Bjorklid E. et al Production and availability of thromboplastin in endothelial cells: the effects of thrombin, endotoxin and platelets. Br J Haematol 1984; 57: 239-46.
  • 9 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol 1989; 134: 1087-97.
  • 10 Brand K, Fowler BJ, Edgington TS. et al Tissue factor mRNA in THP-1 monocytic cells is regulated at both transcriptional and posttranscriptional levels in response to lipopolysaccharide. Mol Cell Biol 1991; 11: 4732-8.
  • 11 Srivastava IK, Liu MA, Gene vaccines. Ann Intern Med 2003; 138: 550-9.
  • 12 Krieg AM, Yi AK, Matson S. et al CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 1995; 374: 546-9.

Correspondence to:

Dr John McVey
Haemostasis and Thrombosis
MRC Clinical Sciences Centre
The Faculty of Medicine, Imperial College
Hammersmith Hospital Campus
Du Cane Road
London, W12 0NN
United Kingdom
Phone: 020 8383 8253   
Fax: 0870 131 3540   

  • References

  • 1 Kemball-Cook G, Johnson DJD, Takamiya O. et al Coagulation factor VII Gln100-->Arg – Amino acid substitution at the epidermal growth factor 2-protease domain interface results in severely reduced tissue factor binding and procoagulant function. J Biol Chem 1998; 273: 8516-21.
  • 2 Dackiw APB, McGilvray ID, Woodside M. et al Prevention of endotoxin-induced mortality by antitissue factor immunization. Arch Surg 1996; 131: 1273-8.
  • 3 Chen D, Giannopoulos K, Shiels PG. et al Inhibition of intravascular thrombosis in murine endotoxemia by targeted expression of hirudin and tissue factor pathway inhibitor analogs to activated endothelium. Blood 2004; 104: 1344-9.
  • 4 Wolff JA, Malone RW, Williams P. et al Direct gene transfer into mouse muscle in vivo. Science 1990; 247: 1465-8.
  • 5 Wilcox JN, Smith KM, Schwartz SM. et al Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl Acad Sci U S A 1989; 86: 2839-43.
  • 6 Archipoff G, Beretz A, Freyssinet JM. et al Heterogeneous regulation of constitutive thrombomodulin or inducible tissue-factor activities on the surface of human saphenous-vein endothelial cells in culture following stimulation by interleukin-1, tumour necrosis factor, thrombin or phorbol ester. Biochem J 1991; 273: 679-84.
  • 7 Galdal KS, Lyberg T, Evensen SA. et al Thrombin induces thromboplastin synthesis in cultured vascular endothelial cells. Thromb Haemost 1985; 54: 373-6.
  • 8 Brox JH, Osterud B, Bjorklid E. et al Production and availability of thromboplastin in endothelial cells: the effects of thrombin, endotoxin and platelets. Br J Haematol 1984; 57: 239-46.
  • 9 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues. Implications for disorders of hemostasis and thrombosis. Am J Pathol 1989; 134: 1087-97.
  • 10 Brand K, Fowler BJ, Edgington TS. et al Tissue factor mRNA in THP-1 monocytic cells is regulated at both transcriptional and posttranscriptional levels in response to lipopolysaccharide. Mol Cell Biol 1991; 11: 4732-8.
  • 11 Srivastava IK, Liu MA, Gene vaccines. Ann Intern Med 2003; 138: 550-9.
  • 12 Krieg AM, Yi AK, Matson S. et al CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 1995; 374: 546-9.