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DOI: 10.1055/s-0037-1614972
Mutations in the αIIb and β3 Genes that Cause Glanzmann Thrombasthenia Can Be Distinguished by a Simple Procedure Using Transformed B-Lymphocytes
Publikationsverlauf
Received
15. Mai 1997
Accepted after resubmission
26. September 1997
Publikationsdatum:
08. Dezember 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199802/lookinside/thumbnails/10-1055-s-0037-1614972-1.jpg)
Summary
Glanzmann thrombasthenia (GT) is caused by a defect in either glycoprotein (GP) IIb (αIIb) or GPIIIa (β3) genes and therefore screening of both genes is required for mutation identification. The β subunit of the GPIIb/IIIa complex (β3) forms a complex with another α subunit (αv) yielding the αvβ3 vitronectin receptor (VnR). GT patients with mutations in the GPIIIa gene that cause diminished synthesis of GPIIIa are deficient in both GPIIb\IIIa and VnR, whereas patients with mutations in the GPIIb gene are deficient in GPIIb\IIIa, yet express normal or increased VnR in their platelets. The presence or absence of VnR in platelet membranes of GT patients has therefore been used for distinguishing between mutations in the GPIIb gene and mutations in the GPIIIa gene. However, the method of assessing VnR in platelets is cumbersome and use of fresh platelets is indispensible. In the present work we devised a procedure for detection of the VnR in B-lymphocytes transformed by Epstein-Bar virus (EBV). The transformed lymphocytes transcribed GPIIIa mRNA but not GPIIb mRNA and expressed VnR on their surface. Using flow cytometry analysis or immuno-precipitation and western blotting VnR was found in B-lymphocytes of GT patients bearing a well characterized mutation in the GPIIb gene. In contrast, in B-lymphocytes of GT patients bearing 2 different mutations in the GPIIIa gene no VnR was detectable. Thus, for determining which gene is mutated in a GT patient, EBV-transformed B-lymphocytes are useful and can as well be used for analyses of GPIIIa mRNA and genomic DNA. Ten ml of blood are sufficient for the procedure.
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References
- 1 Zimrin AB, Eisman R, Vilaire G, Schwartz E, Bennett JS, Poncz M. Structure of platelet glycoprotein IIIa. A common subunit for two different membrane receptors. J Clin Invest 1988; 81: 1470-5.
- 2 Uzan G, Prenant M, Parandinin MH, Martin F, Marguerie G. Tissue specific expression of the platelet GPIIb gene. J Biol Chem 1991; 266: 8932-9.
- 3 Hynes RO. Integrins: Versatility, modulation, and signaling in cell adhesion. Cell 1992; 69: 11-25.
- 4 Coller BS, Cheresh DA, Asch E, Seligsohn U. Platelet vitronectin receptor expression differantiates Iraqi-Jews from Arab patients with Glanzmann thrombasthenia in Israel. Blood 1991; 77: 75-83.
- 5 Peretz H, Rosenberg N, Usher S, Graff E, Newman PJ, Coller BS, Seligsohn U. Glanzmann’s thrombasthenia associated with deletion-insertion and alternative splicing in the glycoprotein IIb gene. Blood 1995; 85: 414-20.
- 6 Yatuv R, Rosenberg N, Dardik R, Brenner B, Seligsohn U. A third mutation causing Glanzmann thrombasthenia in Iraqi-Jews: an A → G transition within the splice site of exon 20 in the GPIIb gene. Thromb Haemost 1997; 1: 361 (abstr).
- 7 Newman PJ, Seligsohn U, Lyman S, Coller BS. The molecular genetic basis of Glanzmann thrombasthenia in the Iraqi-Jewish and Arab population in Israel. Proc Natl Acad Sci USA 1991; 88: 3160-4.
- 8 Rosenberg N, Yatuv R, Orion Y, Zivelin A, Dardik R, Peretz H, Seligsohn U. Glanzmann thrombasthenia caused by an 11.2 kilobase-deletion in the glycoprotein IIIa (β3) is a second mutation in Iraqi-Jews that stemmed from a distinct founder. Blood 1997; 89: 3654-62.
- 9 Coller BS, Seligsohn U, Zivelin A, Zwang E, Lusky A, Modan M. Immunologic and biochemical characterization of homozygous and heterozygous Glanzmann’s thrombasthenia in Iraqi-Jewish and Arab populations in Israel: Comparison of techniques for carrier detection. Br J Haematol 1986; 62: 723-35.
- 10 Miller G, Lipman M. Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc Natl Acad Sci USA 1973; 70: 190-4.
- 11 Peyruchaud O, Nurden A, Bourre F. Efficient RT-PCR on platelet mRNA after long-term storage. Br J Haematol 1997; 96: 183-5.
- 12 Dardik R, Kaufmann Y, Savion N, Rosenberg N, Shenkman B, Varon D. Platelets mediate tumor cell adhesion to the subendothelium under flow conditions: Involvement of platelet GPIIb-IIIa and tumor cell αv integrins. Int J Cancer 1997; 70: 201-7.
- 13 Zimrin AB, Gidwitz S, Lord S, Schwartz E, Bennett JS, White II GC, Poncz M. The genomic organization of platelet glycoprotein IIIa. J Biol Chem 1990; 265: 8590-5.
- 14 Duperray A, Troesch A, Berthiere R, Chagnon E, Frachet P, Uzan G, Margurie G. Biosynthesis and assembly of platelet GPIIb and GPIIIa in human megakaryocytes: Evidence that assembly between pro-GPIIb and GPIIIa is a prerequisite for expression of the complex on the cell surface. Blood 1989; 74: 1603-11.