Journal of Clinical Immunology

, Volume 37, Issue 5, pp 486–495 | Cite as

Multicolor Flow Cytometry for the Diagnosis of Primary Immunodeficiency Diseases

  • Takehiro Takashima
  • Miko Okamura
  • Tzu-wen Yeh
  • Tsubasa Okano
  • Motoi Yamashita
  • Keisuke Tanaka
  • Akihiro Hoshino
  • Noriko Mitsuiki
  • Masatoshi Takagi
  • Eiichi Ishii
  • Kohsuke ImaiEmail author
  • Hirokazu KaneganeEmail author
  • Tomohiro Morio
Original Article



Primary immunodeficiency diseases (PIDDs) are rare inherited diseases that impair the human immune system. We established a multicolor flow cytometric assay to comprehensively evaluate the immune status and immunological characteristics of patients with PIDDs.


Fifty-nine normal controls and 75 patients with PIDDs, including X-linked severe combined immunodeficiency (X-SCID), X-linked agammaglobulinemia (XLA), X-linked hyper IgM syndrome (X-HIGM), ataxia telangiectasia (AT), Wiskott-Aldrich syndrome (WAS), hyper IgE syndrome (HIES), and chronic mucocutaneous candidiasis disease (CMCD), were enrolled in this study. Immunophenotyes were evaluated by multicolor flow cytometry using seven different panels that allowed the detection of major leukocyte populations in peripheral blood.


Multicolor flow cytometry revealed distinct leukocyte populations and immunological features of patients with X-SCID, XLA, X-HIGM, AT, WAS, HIES, and CMCD.


Immunophenotyping by multicolor flow cytometry is useful to evaluate immune status and contributes to the diagnosis and management of patients with PIDDs.


Ataxia telangiectasia Chronic mucocutaneous candidiasis disease Flow cytometry Hyper IgE syndrome Primary immunodeficiency disease 



ataxia telangiectasia


B cell receptor


chronic mucocutaneous candidiasis disease


dendritic cell


double negative T


forward scatter


gain of function


hyper IgE syndrome


invariant NKT


kappa-deleting recombination excision circles


loss of function


myeloid dendritic cells


peripheral blood mononuclear cells


phosphate-buffered saline


plasmacytoid dendritic cells


primary immunodeficiency diseases


recent thymic emigrants


severe combined immunodeficiency


side scatter


T cell receptor


cytotoxic T


follicular helper T


helper T


T cell receptor excision circles


regulatory T


Wiskott-Aldrich syndrome


X-linked hyper IgM syndrome


X-linked agammaglobulinemia


X-linked severe combined immunodeficiency



We thank the patients and their parents as well as the doctors who provided the samples. We thank Ms. Naomi Terada-Takahashi, Ms. Sae Yasuda, Ms. Yuki Inami, Mr. Yohei Kohno, and Mr. Nakaba Ochiai for their technical assistances. This study was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Ministry of Health, Labour, and Welfare of Japan.

Authorship Contribution

Takehiro Takashima, Miko Okamura, Kohsuke Imai, and Hirokazu Kanegane wrote the manuscript. Takehiro Takashima, Miko Okamura, Tzu-Wen Yeh, Tsubasa Okano, Motoi Yamashita, Keisuke Tanaka, and Akihiro Hoshino performed the flow cytometric analysis and collected the data. Noriko Mitsuiki, Masatoshi Takagi, Eiichi Ishii, and Tomohiro Morio contributed to critical discussion. Kohsuke Imai and Hirokazu Kanegane designed the study.

Compliance with Ethical Standards

All subjects or their guardians provided written informed consent to participate in the study in accordance with the Declaration of Helsinki. This study was approved by the Ethics Committee of the Tokyo Medical and Dental University.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Supplementary material

10875_2017_405_MOESM1_ESM.docx (2.6 mb)
ESM 1 (DOCX 2707 kb).


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Takehiro Takashima
    • 1
    • 2
  • Miko Okamura
    • 1
  • Tzu-wen Yeh
    • 1
  • Tsubasa Okano
    • 1
  • Motoi Yamashita
    • 1
  • Keisuke Tanaka
    • 1
  • Akihiro Hoshino
    • 1
    • 3
  • Noriko Mitsuiki
    • 1
  • Masatoshi Takagi
    • 4
  • Eiichi Ishii
    • 2
  • Kohsuke Imai
    • 4
    Email author
  • Hirokazu Kanegane
    • 1
    Email author
  • Tomohiro Morio
    • 1
  1. 1.Department of Pediatrics and Developmental Biology, Graduate School of Medical and Dental SciencesTokyo Medical and Dental University (TMDU)TokyoJapan
  2. 2.Department of PediatricsEhime University Graduate School of MedicineToonJapan
  3. 3.Department of Lifetime Clinical Immunology, Graduate School of Medical and Dental SciencesTokyo Medical and Dental University (TMDU)TokyoJapan
  4. 4.Department of Community Pediatrics, Perinatal and Maternal Medicine, Graduate School of Medical and Dental SciencesTokyo Medical and Dental University (TMDU)TokyoJapan

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