In vitro method establishment for detecting monocyte phagocytosis of sensitized RBCs by flow cytometry
Journal Title: Chinese Journal of Blood Transfusion - Year 2022, Vol 35, Issue 8
Abstract
Objective To establish an experimental method for detecting phagocytosis of sensitized red blood cells in vitro by flow cytometry. Methods Mononuclear cells were isolated from the peripheral blood of blood donors and cultured in a cell incubator for 1 hour, and then adherent monocytes were isolated and obtained. Dib-positive red blood cells (RBCs) were labeled with PKH26 and then sensitized with IgG anti-Dib. The sensitized RBCs were added to monocytes for in vitro phagocytosis assay. Monocytes were labeled with FITC anti-human CD14, then phagocytosis was measured by flow cytometry, and the phagocytic efficiency was calculated. The method was used to detect the phagocytic efficiency of monocytes on human IgG anti-D sensitized RBCs with different titers. Results The phagocytic efficiency of monocytes was averaged at 5% (1.2%~7.6%, SD 3.30) versus 81% (71.4%~92.7%, SD 8.65) in the negative versus positive control group, respectively. Phagocytic activity of monocytes mediated by anti-D was correlated with the antibody titer. The phagocytosis efficiency was within 10% when the antibody titer was lower than 32 and increased sharply when the titer was between 32 to 128, it entered a plateau and stabilized at 80% at the titer above 256. Conclusion A detection platform for detecting phagocytosis-sensitized RBCs in vitro by flow cytometry has been successfully established. It can be used to assess the clinical significance of red blood cell allotype or autologous IgG antibodies.
Authors and Affiliations
Siying ZHU, Yanli JI, Zhen WANG, Dawei CHEN, Zhijian LIAO, Chunyan MO, Jizhi WEN, Guangping LUO, Ling WEI
Keywords
Related Articles
- EP ID EP738315
- DOI 10.13303/j.cjbt.issn.1004-549x.2022.08.008
- Views 30
- Downloads 0
Analysis of platelet apheresis transfusion trends in Suzhou from 2017-2022
[Objective] To investigate the clinical characteristics of patients who underwent multiple platelet transfusions (≥2 times) in Suzhou over the past 5 years (2017-2022), as to provide new insights for patients requiring l...
Analysis on the characteristics of rural blood donation population in Dali Bai Autonomous Prefecture
Objective To explore the status and characteristics of voluntary blood donors in rural areas of Dali Bai Autonomous Prefecture (referred as Dali), and to provide basis for scientific and effective voluntary blood donatio...
Spatial distribution analysis of blood donors′ intended donation addresses based on ArcGIS
Objective To apply the spatial distribution analysis based on ArcGIS software, which has been applied widely in other fields, so as to analyze the intended locations for whole blood donation. Methods After a random strat...
Expert consensus on the resolution level of the methods for HLA genotyping
Human leukocyte antigen (HLA) genotyping has been widely used in establishing the database of the donors for hematopoietic stem cell, HLA matching selection between the donor and recipient, establishing the database of p...
Study on the distribution of ABO blood group in patients with pancreatic cancer
Objective To investigate whether there is a correlation between the differences in ABO blood group distribution in patients with pancreatic cancer, and to evaluate the relative risk. Methods Patients with pathological di...