Simultaneous five cell-lineage flow cytometric analysis system for detection of leucocyte antibodies

Transfusion Medicine
Nobuki MatsuyamaH Shibata

Abstract

Although flow cytometric (FCM) analysis is one of the most widely used approaches to screen the presence of leucocyte antibodies, it has several drawbacks. First, neutrophils and, especially, monocytes exhibit high background reactivity. Second, to determine antibody specificity, it is often necessary to examine not only neutrophils and monocytes but also other lineage cells including T cells, B cells and platelets. Therefore, we attempted to establish an FCM analysis system in which four lineages of leucocytes and platelets are simultaneously tested with low background. FCM analysis was performed using ethylene diamine tetraacetic acid-anticoagulated whole blood as cell sample without any cell preparation. Discrimination of five cell lineages was carried out based on the differences in forward vs. side scatter distribution and in the expression of CD4, CD20 and CD14. When anti-HNA (human neutrophil antigen) 1b antiserum was applied to HNA 1b-positive blood samples, only neutrophils were unambiguously positive. When anti-Naka (anti-CD36) antiserum was applied, only platelets and monocytes were positive. The background reactivity of neutrophils and monocytes was low enough. When anti-human leucocyte antigen (HLA) class II antise...Continue Reading

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Citations

Jan 29, 2011·Hypertension Research : Official Journal of the Japanese Society of Hypertension·Sumihiko HagitaMasayuki Yoshida
Jul 17, 2007·Pediatric Transplantation·Yoshihiro YuiTatsutoshi Nakahata
Jul 25, 2008·Transfusion·Nobuki MatsuyamaHirotoshi Shibata

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