PCR-RFLP Detection od PAI-2 Gene Variants: Prevelence in Ethnic Groups and Disease Relationship in patients Undergoing corony Angiography

Summary

PAI-2 is a fibrinolytic inhibitor produced predominantly by monocytes. Most PAI-2 is intracellular making study in clinical conditions difficult. Abnormalities in production may be associated with inflammation and fibrinolysis at sites of tissue damage such as the atherosclerotic plaque.

PAI-2 gene variants have been described: variant A consists of Asn¹²⁰, Asn⁴⁰⁴ and Ser⁴¹³ and variant B consists of Asp¹²⁰, Lys⁴⁰⁴ and Cys⁴¹³. We designed a PCR-RFLP assay using primers spanning the region containing Asn/Lys⁴⁰⁴ and Ser/Cys⁴¹³. Variant B contains an Mwol restriction site. We analysed 302 Pima Indians and 286 healthy Caucasian volunteers. To investigate relationships between genotype and vascular disease we analysed 333 Caucasian patients undergoing coronary angiography.

Gene variant B was more common in the Pimas than in Caucasians (p <0.0001). There was no significant difference in genotype distribution between the volunteers and patients. In the patients there was no association between genotype and either a history of MI or extent of coronary atheroma.

• References

• 1 Kawano T, Morimoto K, Uemura Y. Urokinase inhibitor in human placenta. Nature 1968; 217: 253-254
• 2 Kruithof EKO, Vassalli JD, Schleuning WD, Mattaliano RJ, Bachmann F. J Biol Chem 1986; 261: 1207-1307
• 3 Kruithof EKO, Tran-Thang C, Gudinchet A, Havert J, Nicoloso G, Genton C, Welti H, Bachmann F. Fibrinolysis in pregnancy. A study of plasminogen activator inhibitors. Blood 1987; 69: 460-469
• 4 Semeraro N, Montemurro P, Giordano P, Santoro N, De Mattia D, Colucci M. Increased mononuclear cell tissue factor and type-2 plasminogen activator inhibitor and reduced plasma fibrinolytic capacity in children with lymphoma. Thromb Haemost 1994; 72: 54-57
• 5 Scherrer A, Kruithof EKO, Grob JP. Plasminogen activator inhibitor-2 in patients with monocytic leukemia. Leukemia 1991; 5: 479-486
• 6 Leung K-C, Byatt JA, Stephens RW. The resistance of fibrin-stimulated tissue plasminogen activator to inactivation by a class PAI-2 inhibitor (miniactivin). Thromb Res 1987; 46: 755-766
• 7 Schleef RR, Wagner NV, Loskutoff DJ. Detection of both type 1 and type 2 plasminogen activator inhibitor. J Cell Physiol 1988; 134: 269-274
• 8 Laug WE, Aebersold R, Jong A, Rideout W, Bergman BL, Baker J. Isolation of multiple types of plasminogen activator inhibitors from vascular smooth muscle cells. Thromb Haemost 1989; 61: 517-521
• 9 Kruithof EKO, Baker MS, Bunn CL. Biological and clinical aspects of plasminogen activator inhibitor type 2. Blood 1995; 86: 4007-4024
• 10 Ritchie H, Jamieson A, Booth NA. Thrombin modulates synthesis of plasminogen activator inhibitor type 2 by human peripheral blood monocytes. Blood 1995; 86: 3428-3435
• 11 Feener E, Northrup J, Aiello L, King G. Angiotensin II induces plasminogen activator inhibitor-1 and -2 expression in vascular endothelial and smooth muscle cells. J Clin Invest 1995; 95: 1353-1362
• 12 Ritchie H, Jamieson A, Booth NA. Peripheral blood monocyte synthesis of plasminogen activator inhibitor 2 in response to native and modified LDL. Thromb Haemost 1995; 74: 1521-1527
• 13 Webb AC, Collins KL, Snyder SE, Alexander SJ, Rosenwasser LJ, Eddy RL, Shows TB, Auron PE. Human monocyte Arg-Serpin cDNA. Sequence, chromosomal assignment, and homology to plasminogen activator-inhibitor. J Exp Med 1987; 166: 77-79
• 14 Antalis TM, Clark MA, Barnes T, Lehrbach PR, Devine PL, Schevzov G, Goss NH, Stephens RW, Tolstoshev P. Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor. Proc Natl Acad Sci USA 1988; 85: 985-999
• 15 Kiso U, Kaudewitz H, Henschen A, Åstedt B, Kruithof EKO, Bachmann F. Determination of intermediates, products and cleavage sites in the reaction between plasminogen activator inhibitor type 2 and urokinase. FEBS Lett 1988; 230: 51-56
• 16 Van den Berg EA, Le Clerq E, Kooistra T, Frants RR, Bakker E. The human gene for plasminogen activator inhibitor 2 (PAI-2) exhibits an EcoRI RFLP. Nucleic Acids Res 1990; 18: 2837
• 17 Young J, Antalis TM, Chenevix-Trench G. Bell RFLP of the plasminogen activator inhibitor type 2 gene (PLANH) on chromosome 18q21-q23. Nucleic Acids Res 1990; 18: 7196
• 18 Ye RD, Wun TC, Sadler JE. cDNA cloning and expression in Escherichia coli of a plasminogen activator inhibitor from human placenta. J Biol Chem 1987; 262: 3718-3725
• 19 Mikus P, Urano T, Lijeström P, Ny T. Plasminogen-activator inhibitor type 2 (PAI-2) is a spontaneously polymerising SERPIN. Biochemical characterisation of the recombinant intracellular and extracellular forms. Eur J Biochem 1993; 218: 1071-1082
• 20 Mansfield MW, Stickland MH, Carter AM, Grant PJ. Polymorphisms of the plasminogen activator inhibitor-1 gene in type 1 and type 2 diabetes, and in patients with diabetic retinopathy. Thromb Haemost 1994; 71: 731-736
• 21 Kruithof EKO, Gudinchet A, Bachmann F. Plasminogen activator inhibitor 1 and plasminogen activator inhibitor 2 in various disease states. Thromb Haemost 1988; 59: 07-12
• 22 Dawson S, Wiman B, Hamsten A, Green F, Humphries S, Henney A. The two allele sequences of a common polymorphism in the promoter of the plasminogen activator inhibitor-1 (PAI-1) gene respond differently to interleukin-1 in Hep G2 cells. J Biol Chem 1993; 268: 10739-10745
• 23 Eriksson P, Kallin B, van’tHooft FM, Båvenholm P, Hamsten A. Allelespecific increase in basal transcription of the plasminogen activator inhibitor-1 gene is associated with myocardial infarction. Proc Natl Acad Sci USA 1995; 92: 1851-1855
• 24 Ossei-Gerning N, Mansfield MW, Stickland MH, Wilson I, Grant PJ. Plasminogen activator inhibitor-1 (PAI-1) promoter 4G/5G genotype and levels in relation to a history of myocardial infarction in patients characterised by coronary angiography. Arterioscl Thromb Vase Biol. 1996 (in press)
• 25 Antalis TM, Godbolt D, Donnan KD, Stringer BW. Southwestern blot mapping of potential regulatory proteins binding to the DNA encoding plasminogen activator inhibitor type 2. Gene 1993; 134: 201-208
• 26 Cousin E, Medcalf RL, Bergonzelli GE, Kruithof EKO. Regulatory elements involved in constitutive and phorbol ester-inducible expression of the plasminogen activator inhibitor type 2 gene promoter. Nucleic Acid Res 1991; 19: 2881-2866
• 27 Dear AE, Medcalf RL. The cellular and molecular biology of plasminogen activator inhibitor type 2. Fibrinolysis 1995; 9: 321-330