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Open Access Highly Accessed Research article

Assessment of Equine Autoimmune Thrombocytopenia (EAT) by flow cytometry

Rafael Nunez14*, M Alice Gomes-Keller3, Colin Schwarzwald2 and Karsten Feige2

Author Affiliations

1 Institutes of Virology, University of Zurich, Zurich, Switzerland

2 Department of Internal Medicine, University of Zurich, Zurich, Switzerland

3 Clinical Laboratory, University of Zurich, Zurich, Switzerland

4 Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, USA

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BMC Blood Disorders 2001, 1:1  doi:10.1186/1471-2326-1-1

Published: 10 April 2001

Abstract

Rationale

Thrombocytopenia is a platelet associated process that occurs in human and animals as result of i) decreased production; ii) increased utilization; iii) increased destruction coupled to the presence of antibodies, within a process know as immune-mediated thrombocytopenia (IMT); or iv) platelet sequestration. Thus, the differentiation of the origin of IMT and the development of reliable diagnostic approaches and methodologies are important in the clarification of IMT pathogenesis. Therefore, there is a growing need in the field for easy to perform assays for assessing platelet morphological characteristics paired with detection of platelet-bound IgG.

Objectives

This study is aimed to develop and characterize a single color flow cytometric assay for detection of platelet-bound IgG in horses, in combination with flow cytometric assessment of platelet morphological characteristics.

Findings

The FSC and SSC evaluation of the platelets obtained from the thrombocytopenic animals shows several distinctive features in comparison to the flow cytometric profile of platelets from healthy animals. The thrombocytopenic animals displayed i) increased number of platelets with high FSC and high SSC, ii) a significant number of those gigantic platelets had strong fluorescent signal (IgG bound), iii) very small platelets or platelet derived microparticles were found significantly enhanced in one of the thrombocytopenic horses, iv) significant numbers of these microplatelet/microparticles/platelet-fragments still carry very high fluorescence.

Conclusions

This study describes the development and characterization of an easy to perform, inexpensive, and noninvasive single color flow cytometric assay for detection of platelet-bound IgG, in combination with flow cytometric assessment of platelet morphological characteristics in horses.