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The Impact of Anticoagulation Agent on the Composition and Phenotype of Blood Leukocytes in Dromedary Camels

Overview
Journal Vet Sci
Publisher MDPI
Date 2022 Feb 24
PMID 35202331
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Abstract

For the analysis of several cellular biomarkers, blood samples are anticoagulated using different agents with different modes of action. However, for the most commonly used anticoagulants, EDTA and heparin, varying effects on blood components have been reported in different species. As little is known about the impact of anticoagulants on the immunological evaluation of camel leukocytes, the present study analyzed the leukogram, the immunophenotype, and the cell vitality of camel leukocytes separated from blood samples anticoagulated with EDTA or lithium heparin. Using flow cytometry and staining with monoclonal antibodies to several cell surface markers, the composition and immunophenotype of camel leukocytes separated from blood anticoagulated with EDTA or heparin were analyzed. In comparison to EDTA-anticoagulated blood, using lithium heparin as an anticoagulant resulted in reduced numbers of total leukocytes and reduced numbers of neutrophils, which led to a reduced neutrophil to lymphocyte ratio. The analysis of cell necrosis and apoptosis after the staining of leukocytes with the DNA-sensitive dye propidium iodide and the mitochondrial membrane potential probe JC1 revealed a higher fraction of necrotic neutrophils and higher fractions of apoptotic neutrophils and monocytes in heparin blood than in EDTA blood. In addition, monocytes from heparin blood showed higher expression levels of the cell surface markers CD14, CD163, and MHCII when compared to cells from EDTA blood. Similarly, in heparin blood, CD44 and CD172a were expressed higher on neutrophils, while CD11a was expressed higher on lymphocytes in comparison to cells from EDTA blood. The results of the current study indicate the importance of considering the type of anticoagulant when investigating the composition, vitality, and immunophenotype of camel leukocytes.

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PMID: 37791077 PMC: 10544901. DOI: 10.3389/fcell.2023.1255823.


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References
1.
el Habbal M, Smith L, Elliott M, Strobel S . Effect of heparin anticoagulation on neutrophil adhesion molecules and release of IL8: C3 is not essential. Cardiovasc Res. 1995; 30(5):676-81. DOI: 10.1016/0008-6363(95)00069-0. View

2.
Nawrocki M, Lory N, Bedke T, Stumme F, Diercks B, Guse A . Trans-Ned 19-Mediated Antagonism of Nicotinic Acid Adenine Nucleotide-Mediated Calcium Signaling Regulates Th17 Cell Plasticity in Mice. Cells. 2021; 10(11). PMC: 8616272. DOI: 10.3390/cells10113039. View

3.
Wolosin J, Zamudio A, Wang Z . Application of JC1 for non-toxic isolation of cells with MDR transporter activity by flow cytometry. PLoS One. 2017; 12(4):e0174905. PMC: 5381900. DOI: 10.1371/journal.pone.0174905. View

4.
OLSEN A, Bladbjerg E, Jensen A, Hansen A . Effect of pre-analytical handling on haematological variables in minipigs. Lab Anim. 2001; 35(2):147-52. DOI: 10.1258/0023677011911516. View

5.
Alan S, Tuna S, Turkoglu E . The relation of neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and mean platelet volume with the presence and severity of Behçet's syndrome. Kaohsiung J Med Sci. 2015; 31(12):626-31. DOI: 10.1016/j.kjms.2015.10.010. View