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Possible application of flow cytometry for evaluation of the structure and functional status of WASP in peripheral blood mononuclear cells
Author(s) -
Nakajima Masaru,
Yamada Masafumi,
Yamaguchi Koji,
Sakiyama Yukio,
Oda Atsushi,
Nelson David L.,
Yawaka Yasutaka,
Ariga Tadashi
Publication year - 2009
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.2008.01180.x
Subject(s) - peripheral blood mononuclear cell , flow cytometry , haematopoiesis , intracellular , wiskott–aldrich syndrome protein , biology , immunology , population , wiskott–aldrich syndrome , peripheral blood , microbiology and biotechnology , cell , stem cell , medicine , genetics , cytoskeleton , gene , in vitro , actin cytoskeleton , environmental health
The Wiskott‐Aldrich syndrome protein (WASP), which is defective in Wiskott‐Aldrich syndrome (WAS) patients, is an intracellular protein expressed in non‐erythroid hematopoietic cells. Previously, we have established methods to detect intracellular WASP expression in peripheral blood mononuclear cells (PBMNCs) using flow cytometric analysis (FCM‐WASP) and have revealed that WAS patients showed absent or very low level intracellular WASP expression in lymphocytes and monocytes, while a significant amount of WASP was detected in those of normal individuals. We applied these methods for diagnostic screening of WAS patients and WAS carriers, as well as to the evaluation of mixed chimera in WAS patients who had previously undergone hematopoietic stem cell transplantation. During these procedures, we have noticed that lymphocytes from normal control individuals showed dual positive peaks, while their monocytes invariably showed a single sharp WASP‐positive peak. To investigate the basis of the dual positive peaks (WASP low‐bright and WASP high‐bright ), we characterized the constituent linage lymphocytes of these two WASP‐positive populations. As a result, we found each WASP low/high population comprised different linage PBMNCs. Furthermore, we propose that the difference between the two WASP‐positive peaks did not result from any difference in WASP expression in the cells, but rather from a difference in the structural and functional status of the WASP protein in the cells. It has been shown that WASP may exist in two forms; an activated or inactivated form. Thus, the structural and functional WASP status or configuration could be evaluated by flow cytometric analysis.