Allergen-specific T-cell response in patients with phenytoin hypersensitivity; simultaneous analysis of proliferation and cytokine production by Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay

Ikuya Tsuge, Akihisa Okumura, Yasuto Kondo, Seiko Itomi, Michiko Kakami, Makiko Kawamura, Yoichi Nakajima, Ryo Komatsubara, Atsuo Urisu

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Background: Phenytoin can induce diversified adverse reactions including generalized eruptions and the hypersensitivity syndrome. Delayed-type allergic mechanisms have been postulated to underlie these reactions. The tests most widely used to detect T-cell sensitization to drugs are the patch test and the lymphocyte transformation test (LTT), but their sensitivity is not sufficient. Simultaneous assessment of both the frequencies and the cytokine-producing phenotypes of allergen-specific T cells has become possible with the recently introduced carboxyfluorescein succinimidyl ester (CFSE) assay. Methods: Seven patients who presented with phenytoin-induced maculopapular exanthema with and without fever were included in this study. Peripheral blood mononuclear cells (PBMCs) were labeled with CFSE and cultured with phenytoin for seven days. The cells were stained with anti-CD4 and cytokine-specific monoclonal antibodies (MoAbs), and analyzed with FACSCalibur. Results: The phenytoin-specific proliferation of CD4+ cells in patients was significantly higher than in the four controls exposed to phenytoin, and in seven healthy children with no previous phenytoin intake. A significant difference in the percentages of CD4+ IFN-γ+ cells between patients and the seven healthy children was observed. The sensitivity and specificity of proliferation were 100% and 90.9%, and those of IFN-γ secretion were 71.4% and 100%, respectively. Conclusions: Phenytoin-specific proliferation may be detected with greater sensitivity by the CFSE dilution assay than the conventional LTT. The assay revealed that both CD4+ and CD4- T cells proliferated and produced IFN-γ and TNF-α after stimulation with phenytoin. The CFSE dilution assay might be useful for the diagnosis and understanding of drug hypersensitivity.

Original languageEnglish
Pages (from-to)149-155
Number of pages7
JournalAllergology International
Volume56
Issue number2
DOIs
Publication statusPublished - 01-01-2007

Fingerprint

Phenytoin
Allergens
Hypersensitivity
Esters
Cytokines
T-Lymphocytes
Lymphocyte Activation
Drug Hypersensitivity
Patch Tests
6-carboxyfluorescein
Exanthema
Blood Cells
Fever
Monoclonal Antibodies
Cell Proliferation
Phenotype
Sensitivity and Specificity
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy

Cite this

Tsuge, Ikuya ; Okumura, Akihisa ; Kondo, Yasuto ; Itomi, Seiko ; Kakami, Michiko ; Kawamura, Makiko ; Nakajima, Yoichi ; Komatsubara, Ryo ; Urisu, Atsuo. / Allergen-specific T-cell response in patients with phenytoin hypersensitivity; simultaneous analysis of proliferation and cytokine production by Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay. In: Allergology International. 2007 ; Vol. 56, No. 2. pp. 149-155.
@article{eb2dd5703dcf417f9f1ab7f40832dc8a,
title = "Allergen-specific T-cell response in patients with phenytoin hypersensitivity; simultaneous analysis of proliferation and cytokine production by Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay",
abstract = "Background: Phenytoin can induce diversified adverse reactions including generalized eruptions and the hypersensitivity syndrome. Delayed-type allergic mechanisms have been postulated to underlie these reactions. The tests most widely used to detect T-cell sensitization to drugs are the patch test and the lymphocyte transformation test (LTT), but their sensitivity is not sufficient. Simultaneous assessment of both the frequencies and the cytokine-producing phenotypes of allergen-specific T cells has become possible with the recently introduced carboxyfluorescein succinimidyl ester (CFSE) assay. Methods: Seven patients who presented with phenytoin-induced maculopapular exanthema with and without fever were included in this study. Peripheral blood mononuclear cells (PBMCs) were labeled with CFSE and cultured with phenytoin for seven days. The cells were stained with anti-CD4 and cytokine-specific monoclonal antibodies (MoAbs), and analyzed with FACSCalibur. Results: The phenytoin-specific proliferation of CD4+ cells in patients was significantly higher than in the four controls exposed to phenytoin, and in seven healthy children with no previous phenytoin intake. A significant difference in the percentages of CD4+ IFN-γ+ cells between patients and the seven healthy children was observed. The sensitivity and specificity of proliferation were 100{\%} and 90.9{\%}, and those of IFN-γ secretion were 71.4{\%} and 100{\%}, respectively. Conclusions: Phenytoin-specific proliferation may be detected with greater sensitivity by the CFSE dilution assay than the conventional LTT. The assay revealed that both CD4+ and CD4- T cells proliferated and produced IFN-γ and TNF-α after stimulation with phenytoin. The CFSE dilution assay might be useful for the diagnosis and understanding of drug hypersensitivity.",
author = "Ikuya Tsuge and Akihisa Okumura and Yasuto Kondo and Seiko Itomi and Michiko Kakami and Makiko Kawamura and Yoichi Nakajima and Ryo Komatsubara and Atsuo Urisu",
year = "2007",
month = "1",
day = "1",
doi = "10.2332/allergolint.O-06-457",
language = "English",
volume = "56",
pages = "149--155",
journal = "Allergology International",
issn = "1323-8930",
publisher = "Japanese Society of Allergology",
number = "2",

}

Allergen-specific T-cell response in patients with phenytoin hypersensitivity; simultaneous analysis of proliferation and cytokine production by Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay. / Tsuge, Ikuya; Okumura, Akihisa; Kondo, Yasuto; Itomi, Seiko; Kakami, Michiko; Kawamura, Makiko; Nakajima, Yoichi; Komatsubara, Ryo; Urisu, Atsuo.

In: Allergology International, Vol. 56, No. 2, 01.01.2007, p. 149-155.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Allergen-specific T-cell response in patients with phenytoin hypersensitivity; simultaneous analysis of proliferation and cytokine production by Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay

AU - Tsuge, Ikuya

AU - Okumura, Akihisa

AU - Kondo, Yasuto

AU - Itomi, Seiko

AU - Kakami, Michiko

AU - Kawamura, Makiko

AU - Nakajima, Yoichi

AU - Komatsubara, Ryo

AU - Urisu, Atsuo

PY - 2007/1/1

Y1 - 2007/1/1

N2 - Background: Phenytoin can induce diversified adverse reactions including generalized eruptions and the hypersensitivity syndrome. Delayed-type allergic mechanisms have been postulated to underlie these reactions. The tests most widely used to detect T-cell sensitization to drugs are the patch test and the lymphocyte transformation test (LTT), but their sensitivity is not sufficient. Simultaneous assessment of both the frequencies and the cytokine-producing phenotypes of allergen-specific T cells has become possible with the recently introduced carboxyfluorescein succinimidyl ester (CFSE) assay. Methods: Seven patients who presented with phenytoin-induced maculopapular exanthema with and without fever were included in this study. Peripheral blood mononuclear cells (PBMCs) were labeled with CFSE and cultured with phenytoin for seven days. The cells were stained with anti-CD4 and cytokine-specific monoclonal antibodies (MoAbs), and analyzed with FACSCalibur. Results: The phenytoin-specific proliferation of CD4+ cells in patients was significantly higher than in the four controls exposed to phenytoin, and in seven healthy children with no previous phenytoin intake. A significant difference in the percentages of CD4+ IFN-γ+ cells between patients and the seven healthy children was observed. The sensitivity and specificity of proliferation were 100% and 90.9%, and those of IFN-γ secretion were 71.4% and 100%, respectively. Conclusions: Phenytoin-specific proliferation may be detected with greater sensitivity by the CFSE dilution assay than the conventional LTT. The assay revealed that both CD4+ and CD4- T cells proliferated and produced IFN-γ and TNF-α after stimulation with phenytoin. The CFSE dilution assay might be useful for the diagnosis and understanding of drug hypersensitivity.

AB - Background: Phenytoin can induce diversified adverse reactions including generalized eruptions and the hypersensitivity syndrome. Delayed-type allergic mechanisms have been postulated to underlie these reactions. The tests most widely used to detect T-cell sensitization to drugs are the patch test and the lymphocyte transformation test (LTT), but their sensitivity is not sufficient. Simultaneous assessment of both the frequencies and the cytokine-producing phenotypes of allergen-specific T cells has become possible with the recently introduced carboxyfluorescein succinimidyl ester (CFSE) assay. Methods: Seven patients who presented with phenytoin-induced maculopapular exanthema with and without fever were included in this study. Peripheral blood mononuclear cells (PBMCs) were labeled with CFSE and cultured with phenytoin for seven days. The cells were stained with anti-CD4 and cytokine-specific monoclonal antibodies (MoAbs), and analyzed with FACSCalibur. Results: The phenytoin-specific proliferation of CD4+ cells in patients was significantly higher than in the four controls exposed to phenytoin, and in seven healthy children with no previous phenytoin intake. A significant difference in the percentages of CD4+ IFN-γ+ cells between patients and the seven healthy children was observed. The sensitivity and specificity of proliferation were 100% and 90.9%, and those of IFN-γ secretion were 71.4% and 100%, respectively. Conclusions: Phenytoin-specific proliferation may be detected with greater sensitivity by the CFSE dilution assay than the conventional LTT. The assay revealed that both CD4+ and CD4- T cells proliferated and produced IFN-γ and TNF-α after stimulation with phenytoin. The CFSE dilution assay might be useful for the diagnosis and understanding of drug hypersensitivity.

UR - http://www.scopus.com/inward/record.url?scp=34250840027&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34250840027&partnerID=8YFLogxK

U2 - 10.2332/allergolint.O-06-457

DO - 10.2332/allergolint.O-06-457

M3 - Article

C2 - 17460442

AN - SCOPUS:34250840027

VL - 56

SP - 149

EP - 155

JO - Allergology International

JF - Allergology International

SN - 1323-8930

IS - 2

ER -