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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

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1437-4331
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Volume 53, Issue 1

Issues

Quantification of adult T-cell leukemia/lymphoma cells using simple four-color flow cytometry

Tomohiro Ishigaki
  • Corresponding author
  • Clinical FACS Core Laboratory, Institute of Medical Science, University of Tokyo, Tokyo, Japan
  • Institute of Medical Science, Division of Stem Cell Therapy, University of Tokyo, Tokyo, Japan
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/ Yuji Zaike
  • Institute of Medical Science, Department of Laboratory Medicine, Research Hospital, University of Tokyo, Tokyo, Japan
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/ Masanori Nojima
  • Institute of Medical Science, Division of Advanced Medicine Promotion, University of Tokyo, Tokyo, Japan
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/ Seiichiro Kobayashi
  • Institute of Medical Science, Division of Molecular Therapy, University of Tokyo, Tokyo, Japan
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/ Nobuhiro Ohno
  • Institute of Medical Science, Department of Hematology and Oncology, Research Hospital, University of Tokyo, Tokyo, Japan
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/ Kaoru Uchimaru
  • Institute of Medical Science, Department of Hematology and Oncology, Research Hospital, University of Tokyo, Tokyo, Japan
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/ Arinobu Tojo
  • Institute of Medical Science, Division of Molecular Therapy, University of Tokyo, Tokyo, Japan
  • Institute of Medical Science, Department of Hematology and Oncology, Research Hospital, University of Tokyo, Tokyo, Japan
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/ Hiromitsu Nakauchi
  • Institute of Medical Science, Division of Stem Cell Therapy, University of Tokyo, Tokyo, Japan
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/ Nobukazu Watanabe
  • Clinical FACS Core Laboratory, Institute of Medical Science, University of Tokyo, Tokyo, Japan
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Published Online: 2014-07-23 | DOI: https://doi.org/10.1515/cclm-2014-0183

Abstract

Background: The absolute number of adult T-cell leukemia/lymphoma (ATL) cells in peripheral blood is an essential indicator to evaluate disease status. However, microscopically counting ATL cells based on morphology requires experience and tends to be inaccurate due to the rarity of ATL.

Methods: Based on our research showing that acute-type ATL cells are specifically enriched in the CD4+/CD7- (CD7N) fraction, a new analytical method to accurately quantify ATL cells was established using an internal bead standard and simple four-color flow cytometry. This method was verified by comparison with microscopic examination of 49 peripheral blood samples and used to follow up patients.

Results: A strong correlation was observed between the number of CD7N cells measured by flow cytometry and the number of abnormal lymphocytes measured microscopically by experienced technicians [Pearson’s R, 0.963; Spearman’s rho, 0.921; intercorrelation coefficient, 0.962]. The linear regression coefficient was close to 1 (β=1.013). Our method could detect 1 cell/μL, and the limit of quantitation was between 2.9 and 9.8 cells/μL. The frequency of CD7N cells among CD4+ cells changed during chemotherapy, which reflected differences between chemosensitive and chemoresistant cases. Kaplan-Meier analysis with a log-rank test showed that patients with decreased CD7N proportion after chemotherapy had significantly longer disease-specific survival (p=0.003).

Conclusions: Our newly established method quantified tumor cells in patients with acute-type ATL. Furthermore, this method was useful for assessing the efficacy of chemotherapy, and the change of the CD7N proportion could be more important to predict prognosis.

This article offers supplementary material which is provided at the end of the article.

Keywords: adult T-cell leukemia/lymphoma (ATL); flow cytometry; HAS-Flow; human T-cell leukemia virus type 1 (HTLV-1)

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About the article

Corresponding author: Tomohiro Ishigaki, Clinical FACS Core Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan, Phone: +81 3 5449 5765, Fax: +81 3 5449 5750, E-mail: ; Institute of Medical Science, Division of Stem Cell Therapy, University of Tokyo, Tokyo, Japan


Received: 2014-02-19

Accepted: 2014-06-18

Published Online: 2014-07-23

Published in Print: 2015-01-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 53, Issue 1, Pages 85–93, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2014-0183.

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