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Pregled bibliografske jedinice broj: 167128

Zbornik radova

Autori: Vidović, Anđelko; Gotovac, Katja; Vilibić, Maja; Sabioncello, Ante; Rabatić, Sabina; Folnegović-Šmalc, Vera; Markotić, Alemka; Dekaris, Dragan
Naslov: Flow cytometry monitoring of leukocyte-platelet aggregates in PTSD
( Flow cytometry monitoring of leukocyte-platelet aggregates in PTSD )
Izvornik: Croatian Immunological Society, Annual Meeting 2004 / Jonjić, Stipan (ur.). - Opatija :
Skup: Annual Meeting of the Croatian Immunological Society 2004
Mjesto i datum: Opatija, Hrvatska, 8-10.10. 2004
Ključne riječi: platelet; leukocyte; aggregates; flow cytometry; PTSD; stress; cardiovascular disease
( platelet; leukocyte; aggregates; flow cytometry; PTSD; stress; cardiovascular disease )
Sažetak:
There is a growing evidence for influence of psychological factors on development of coronary artery disease (CAD). It is proposed that platelet-mediated mechanisms might form one of the pathways through which this association is formed. Whole blood flow cytometry is a valuable method for assessing platelet function and measurement of circulating leukocyte-platelet aggregates (LPA) provides a sensitive marker for in vivo platelet activation. We have recently introduced this method in our laboratory and our preliminary study obtained on 14 PTSD patients and 8 aged matched healthy controls showed reproducible results comparable with recent literature. We assume that increased leukocyte-platelet aggregation may frequently be induced in chronic stress and may accumulate to confer increased cardiovascular disease risk on susceptible individuals. A clean blood draw and gentle handling of specimens are required to avoid spontaneous platelet activation. Samples were drawn using 20 gauge needles in Citrate theophylline adenosine dipyridimole (CTAD) Vacutainers which chelates Ca2+ and increases intracellular cAMP, keeping platelets "quiet". 10 micro litre of blood was immediately added to 90 micro litre solution containing CD45 FITC, CD14 PE (Leucogate) and CD42a PerCP (platelet specific antibody) monoclonal antibodies. After 20 minutes of incubation at room temperature in the dark, samples were fixed using 900 micro litre of 0.5% paraformaldehyde and stored at 4 degree C for at least 2 hours. We analyzed samples with Becton Dickinson LSR II flow cytometer using CD45 and FSC threshold gating to get rid of unwanted events in unlysed and unwashed whole blood. Monocytes, neutrophils and lymphocytes could be distinguished based on linear forward versus side light-scatter properties, but identification of gated populations was improved by using a leukocyte-specific antibodies. Proportions of CD42a positive events in monocyte, neutrophil and lymphocyte gates were measured. Our results did not reveal any significant difference between PTSD patients and healthy controls although percentages of monocytes platelets aggregates tend to be higher in PTSD patients. This may be due to small sample sizes so further investigation is needed. Furthermore, better choice of monoclonal antibodies and multiparameter capabilities of LSR II flow cytometer might improve our detection of LPA and reveal possible risk for CAD in people under prolonged stress.
Vrsta sudjelovanja: Poster
Vrsta prezentacije u zborniku: Sažetak
Vrsta recenzije: Domaća recenzija
Projekt / tema: 0021003
Izvorni jezik: eng
Kategorija: Znanstveni
Znanstvena područja:
Temeljne medicinske znanosti,Kliničke medicinske znanosti
Puni text rada: 167128.Abstracts_-_HID2004_andelko_trombociti.doc



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