A novel multiparameter flow cytometric assay for inosine triphosphatase expression analysis in leukocytes

Wim H. M. Vroemen, Imke C. A. Munnix, Jaap A. Bakker, Jorgen Bierau, Mark Huts, Mathie P. G. Leers*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The aim of this study was to assess inosine triphosphate (ITPase) expression in the different leukocyte populations present in peripheral blood samples of a nonimmune compromised control group. For this purpose, a multiparameter flow cytometric assay was developed and performed to study ITPase expression in peripheral leukocyte subpopulations of healthy volunteers (n = 20). Qualitative ITPase expression was assessed by determining the percentage of ITPase-positive cells. Quantitative data were obtained by measuring the median fluorescent intensity (MFI). Subcellular localization of ITPase was analyzed using immunocytochemistry. Immunocytochemistry showed that ITPase is present in all leukocytes and localized intracellular. Based on this finding, a multiparameter flow cytometric assay was developed using a Fix & Perm strategy. Qualitative and quantitative ITPase expression remained stable (variation,
Original languageEnglish
Pages (from-to)672-678
JournalCytometry Part A
Volume81A
Issue number8
DOIs
Publication statusPublished - Aug 2012

Keywords

  • ITPase
  • flow cytometry
  • leukocyte subpopulations
  • thiopurines

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