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Hematological Cellular Alterations in Plateletpheresis Donors

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Hematological Cellular Alterations in Plateletpheresis Donors Int J Blood Transfus Immunohematol 2018;8:100041Z02HF2018. Fayed et al. 1 www.ijbti.com ORIGINAL ARTICLE PEER REVIEWED OPEN| OPEN ACCESS ACCESS Hematological cellular alterations in plateletpheresis donors Hanan Mahmoud Fayed, Sanaa Shaker Ali, Eman Salah Eldin ABSTRACT count (p = 0.041) with an insignificant increase in absolute monocyte (p = 0.103), lymphocyte Aims: Blood contact with foreign surfaces in the subsets count, CD4:CD8 ratio, and WBC count. apheresis systems may activate many cell types However, there was significant increase in the that return back to the donor. However, the median platelets complexed with neutrophil, safety issue regarding post-procedure platelet lymphocytes, and monocytes. Conclusion: activation is not well assessed. We aimed All donors had a significant drop in all blood to evaluate the alterations in hematological counts; none of them manifested features parameters and to explore the formation of of thrombocytopenia or anemia. However, platelets leukocytes aggregates (PLAs) and/or an increase in PLAs formation provides an complexes in healthy donors who underwent evidence of ongoing platelet activation, a first-time plateletpheresis procedure. Methods: platelet-leucocyte interaction that may induce a Blood from 100 healthy donors were assessed pre-thrombotic risk. This result is important to by BD FACS Calibur flow cytometer for: a) consider as it might have potential therapeutic detection of PLAs using (CD41, CD42b, CD61) implications. Nevertheless, more prospective antibodies against platelets surface molecules studies are essential to establish guidelines for to detect its expression on neutrophils, donor safety. monocytes, lymphocytes, and b) evaluation of red cell mechanical fragility (RBC-MF). Results: Keywords: Donor safety, Flow cytometry, Plate- After donation a significant decrement of donor let-leukocyte aggregates, Single donor platelet blood cell counts; the percent (%) reduction in hemoglobin (Hb) 7.9(5.1–9.2) (p = 0.017), How to cite this article hematocrit (Hct) 6(2.72-7.81) (p = 0.043) %, residual red cells (p = 0.016), platelet count Fayed HM, Ali SS, Eldin ES. Hematological cellular (PLT) 22.7(9.5–32) (p = 0.031) together with a alterations in plateletpheresis donors. Int J Blood significant increase in the MPV (p = 0.001), the Transfus Immunohematol 2018;8:100041Z02HF2018. absolute neutrophil (p = 0.026) and lymphocyte ********* Hanan Mahmoud Fayed1, Sanaa Shaker Ali2, Eman Salah Article ID: 100041Z02HF2018 Eldin3 Affiliations: 1Assistant Professor and Charge d’affaires, Clinical and Chemical Pathology Department, Faculty of ********* Medicine-South Valley University, Qena, Egypt; 2Assistant doi: 10.5348/100041Z02HF2018OA Professor, Clinical and Chemical Pathology Department, Faculty of Medicine-South Valley University, Qena, Egypt; 3Lecturer, Clinical and Chemical Pathology Department, Faculty of Medicine-Assiut University, Assiut-Egypt. Corresponding Author: Hanan Mahmoud Fayed, Qena Fac- INTRODUCTION ulty of Medicine South Valley University 6th km Qena Safa- ga, Road, Qena, Egypt 82523; Email: hananfayed@yahoo. The rapidly expanded clinical uses of blood component com; [email protected] therapy resulted in great improvement in the field of therapeutic apheresis [1]. The concerns of patient care are given highest priority, Received: 02 March 2018 therefore substantial efforts were made to support the Accepted: 03 May 2018 widespread use of single donor platelet-apheresis (SDP) Published: 13 June 2018 instead of platelet concentrate (PC), and these are mainly International Journal of Blood transfusion and Immunohematology, Vol. 8; 2018. ISSN: 2230-9020 Int J Blood Transfus Immunohematol 2018;8:100041Z02HF2018. Fayed et al. 2 www.ijbti.com relevant to the care of patients with hematologic diseases Blood samples [2]. Moreover; SDP has many advantages including Five milliliter venipuncture blood sample was economic use of blood due to selective components drawn twice from the donors; (before the beginning collection with the elimination of unnecessary component of the apheresis procedure and after the completion of separation in the laboratory, the possibility of frequent the apheresis procedure), from a vein that was not to donations, reduced donor exposures and hence reduced be used during the procedure), with minimum stasis risk of alloimmunization [3]. through a 21-gauge needle into sterile evacuated tubes Platelets apheresis procedures are usually safe and (BD Vacutainer, San Jose, CA) containing EDTA and well tolerated. However, it can cause adverse local or 3.8% sodium citrate with a blood: anticoagulation ratio systemic reactions to the donors [4]. of 9:1) (to reduce art factual platelets activation due to Apheresis may cause alterations in donor’s tissue factor contamination, the first blood sample drawn hemorheology because of anti-coagulant used, changes was not used for flow cytometer (FCM)). All samples in blood composition and extracorporeal circulation [5]. were processed within 10 min of collection for complete We aimed to evaluate the alterations in cellular blood count (CBC), assessment of RBCs osmotic fragility hematological parameters and to explore the formation of assessment besides platelets activation and formation of platelets leukocytes aggregates (PLAs) [platelet-monocyte platelet-leukocyte aggregates (PLAs) in blood [platelet- complexes (PMC), platelets-neutrophil complexes (PNC), monocyte complex (PMC), platelet-neutrophil complex and platelets-lymphocyte complexes (PLC)] in healthy (PNC) and platelet-lymphocyte complex (PLC)] were donors who underwent first-time platelet-apheresis determined by FACS Calibur FCM (BD Biosciences, San procedure. Jose, CA, USA); after proper setting, calibration, and compensation; data acquisition and analyzes was done by Cell Quest Pro software (v5.2). MATERIALS AND METHODS • CBC by a calibrated hematology analyzer Cell- Dyn 3500 (Abbott Diagnostics, IL-USA). Donor Study population platelet percentage (%) loss was calculated using the following formula: platelet loss = (pre-platelet A cross-sectional study involving hundred healthy count) – (post-platelet count) × 100 / (pre-platelet first-time platelet-apheresis donors selected with the count). The percent change of donor MPV = the following criteria: weight > 60 kg; age between 18 and difference in the donor’s pre- and post-procedure 50 years; hemoglobin > 12.5 gm/dl and platelets count > MPV × 100 / (pre-MPV). 200x109/L; negative serological tests for HIV, hepatitis • Lymphocyte subsets: Using whole blood B surface antigen & hepatitis C and syphilis; absence of samples; 2 ml blood in EDTA vacutainer tube for any illness; none of them used anti-inflammatory drugs CBC using cell dyne-1800 (Abbott Diagnostics, and with suitable venous accesses. All donations were USA) and analysis of peripheral blood lymphocyte performed at a tertiary care blood transfusion center using BD FACS Caliber™; (Becton Dickinson of Assiut University Hospitals from March 2017 to Biosciences, USA). T cell subset performed via December 2017. The study was conducted in accordance tricolor using the following fluorescent-labeled with the Helsinki Declaration as revised in 2008. monoclonal antibodies from Becton Dickinson Procedure details were explained to each donor who gave (BD, San Jose, CA-USA); (CD3 [peridinin- informed consent before the procedure. Ante-cubital chlorophyl proteins (PerCP)], CD4 [fluorescent veins were used for the venipuncture in all the donors. isothyocyanate (FITC)], and CD8 [phycoerythrin Vital signs were monitored at the beginning and end of (PE)] as described by the manufacturer. Data each procedure; donors were also monitored for adverse processing was carried out using Cell quest 3.0.1 events during the procedures. software. Acquisition of cells was standardized for 10,000 events per sample and fluorescence was Single-donor platelet equipment measured on a logarithmic scale. A lymphocytes gate was created based on their forward and Platelet apheresis was obtained using Fenwal side scatter properties, i.e., size and granularity; Amicus™ separator version 3.1 software; single- the percentages and the absolute count of the needle system platelet-apheresis kit (PL 2410, Fenwal lymphocyte subpopulations were calculated Inc.) (Fenwal, Lake Zurich, IL, USA) used as per the by multiplying the percentage (results of flow manufacturer’s standard operating procedure with a cytometer) by absolute lymphocyte counts whole blood (WB)/acid citrate dextrose (ACD-A) ratio (results of CBC), (Figure 1). of 11:1, interface set point 0.60 and blood flow rate 60– • Flow-cytometry Osmotic Fragility Test 75 ml/ min. The donors’ data (weight, sex, height, Hb, (FCM-OF): Done according to the process and pre-apheresis PB PLT count) were entered into the described by (Won and Suh) with a hemolysis cell separator program to determine the blood volume inducing agent, distilled water (DW), was spiked processed to reach the target platelets yield (3×1011). International Journal of Blood transfusion and Immunohematology, Vol. 8; 2018. ISSN: 2230-9020 Int J Blood Transfus Immunohematol 2018;8:100041Z02HF2018. Fayed et al. 3 www.ijbti.com Figure 2: (A and B): Red cell fragility before and after platelets-apheresis procedure: (A) The FSC/time plot of red cells [% residual red cells before and after plateletapheresis]; (B) The FSC/time plot of red cells [% residual red cells after Figure 1: Dot plot analysis of peripheral blood lymphocyte plateletapheresis]. subsets: (A) Lymphocyte
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