Assessment of Equine Autoimmune Thrombocytopenia (EAT) by flow cytometry.
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.
Publication Date: 2001-04-21 PubMed ID: 11313001PubMed Central: PMC32162DOI: 10.1186/1471-2326-1-1Google Scholar: Lookup
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- Journal Article
Summary
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This research aims to develop a simple and cost-effective test to detect immune-related platelet issues in horses, using something called a flow cytometric assay.
Background and Rationale
- Thrombocytopenia is a condition that can occur in both humans and animals, characterized by abnormal platelet counts.
- This abnormality can be due to different factors: reduced platelet production, increased utilization, excessive destruction linked with the presence of antibodies (a process known as immune-mediated thrombocytopenia or IMT), or platelet sequestration.
- Understanding the root cause of IMT and the development of accurate diagnostic methods is vital to clarify how it develops. The research field, thus, requires accessible assays to assess platelet attributes along with the ability to detect platelet-bound Immunoglobulin G (IgG), an antibody that reacts with the body’s own cells or tissues.
Objectives of the Research
- The main goal of this research was to create and evaluate a flow cytometric assay (a procedure used to measure the physical and chemical characteristics of a population of cells or particles) for the detection of platelet-bound IgG in horses.
- The study also aimed to pair this process with the evaluation of platelet morphological characteristics via flow cytometry, a method that analyses cells based on their size, granularity, and fluorescence.
Key Findings of the Study
- The study observed several specific features when comparing platelets from healthy and thrombocytopenic animals.
- The thrombocytopenic animals showed an increase in platelets with high forward scatter (FSC) and high side scatter (SSC), indicators of cell size and complexity respectively.
- A significant number of these larger platelets had a strong fluorescence signal, suggesting they had bound IgG.
- Also, one thrombocytopenic horse displayed a significant increase in very small platelets or platelet-derived microparticles. Many of these cell fragments also had a high fluorescence, indicating the presence of IgG.
Conclusions
- The research successfully developed and evaluated a straightforward, affordable, and noninvasive single-color flow cytometric assay to detect platelet-bound IgG.
- Alongside this, the study also assessed the physical characteristics of the platelets in horses, contributing to the understanding of equine autoimmune thrombocytopenia (EAT), and potentially paving the way for better diagnostics and treatment approaches in the future.
Cite This Article
APA
Nunez R, Gomes-Keller MA, Schwarzwald C, Feige K.
(2001).
Assessment of Equine Autoimmune Thrombocytopenia (EAT) by flow cytometry.
BMC Blood Disord, 1(1), 1.
https://doi.org/10.1186/1471-2326-1-1 Publication
Researcher Affiliations
- Institutes of Virology, University of Zurich, Zurich, Switzerland. nunezr@mskcc.org
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