Bergmeier, Wolfgang and Schulte, Valerie and Brockhoff, Gero and Bier, Ulrich and Zirngibl, Hubert and Nieswandt, Bernhard (2002) Flow cytometric detection of activated mouse integrin aIIb beta 3 with a novel monoclonal antibody. CYTOMETRY, 48 (2). pp. 80-86. ISSN 0196-4763
Full text not available from this repository. (Request a copy)Abstract
Background: Integrin alphaIIbbeta3 mediates platelet adhesion and aggregation and plays a crucial role in thrombosis and hemostasis. alphaIIbbeta3 is expressed in a low affinity state on resting platelets. Upon platelet activation, alphaIIbbeta3 shifts to a high affinity conformation that efficiently binds its ligands. On human platelets, the high affinity conformation of alphaIIbbeta3 is detected by the monoclonal antibody (mAb), PAC-1. However, a reagent with binding specificity to high affinity mouse alphaIIbbeta3 has not been described so far. Methods: A novel rat mAb directed against mouse alphaIIbbeta3 (JON/A) was generated and characterized. JON/A was conjugated with fluorescein isothiocyanate (JON/A(FITC)) or with R-phycoerythrin (JON/A(PE)) and used for flow cytometric analysis of mouse platelets. Results: Although JON/AF(ITC) bound to resting and activated platelets, virtually no binding of the larger JON/A(PE) to resting platelets was detectable. However, strong binding of JON/A(PE) occurred on platelet activation in a dose-dependent manner. Binding of JON/A(PE) required extracellular free calcium and was irreversible, thereby stabilizing the high affinity conformation of alphaIIbbeta3. Conclusion: JON/A(PE) is the first tool for direct assessment of integrin alphaIIbbeta3 activation in mice. Furthermore, JON/A(FITC) and JON/A(PE) provide the first examples of fluorescent antibody derivatives with identical antigenic specificitiy that allow the discrimination between the resting and the activated state of an integrin.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | GLYCOPROTEIN-IIB-IIIA; PLATELET ACTIVATION; GPIIB-IIIA; MICE; COMPLEX; ALPHA-IIB-BETA-3; RECEPTORS; ANTIGEN; MODEL; POOL; flow cytometry; mouse; platelet; integrin aIIb beta 3; activation |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 25 Oct 2021 11:07 |
| Last Modified: | 25 Oct 2021 11:07 |
| URI: | https://pred.uni-regensburg.de/id/eprint/40182 |
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