Quantification of adult T-cell leukemia/lymphoma cells using simple four-color flow cytometry
-
Tomohiro Ishigaki
,
Yuji Zaike
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.
Acknowledgments
We would like to thank Dr. Naoki Oyaizu and Dr. Naoyuki Isoo (Laboratory Medicine, Research Hospital, Institute of Medical science) for their kind permission to introduce this clinical test, as well as Mr. Yukihisa Tanaka, Ms. Etsuko Nagai, and Ms. Motoko Mizukami for their excellent morphological classification.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Financial support: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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Supplemental Material
The online version of this article (DOI: DOI:10.1515/cclm-2014-0183) offers supplementary material, available to authorized users.
©2015 by De Gruyter
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- Harmonisation of specialist training and continuing professional development in laboratory medicine: a long but necessary journey
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