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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

Abstract

Purpose

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.

Methods

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.

Results

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

Conclusions

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

Keywords

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

Abbreviations

AT

ataxia telangiectasia

BCR

B cell receptor

CMCD

chronic mucocutaneous candidiasis disease

DC

dendritic cell

DNT

double negative T

FSC

forward scatter

GOF

gain of function

HIES

hyper IgE syndrome

iNKT

invariant NKT

KRECs

kappa-deleting recombination excision circles

LOF

loss of function

mDCs

myeloid dendritic cells

PBMCs

peripheral blood mononuclear cells

PBS

phosphate-buffered saline

pDCs

plasmacytoid dendritic cells

PIDDs

primary immunodeficiency diseases

RTEs

recent thymic emigrants

SCID

severe combined immunodeficiency

SSC

side scatter

TCR

T cell receptor

Tc

cytotoxic T

TFH

follicular helper T

Th

helper T

TRECs

T cell receptor excision circles

Treg

regulatory T

WAS

Wiskott-Aldrich syndrome

X-HIGM

X-linked hyper IgM syndrome

XLA

X-linked agammaglobulinemia

X-SCID

X-linked severe combined immunodeficiency

Notes

Acknowledgements

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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