Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3

Kazuo Yamawaki, Chisato Inuo, Takayasu Nomura, Kenichi Tanaka, Yoichi Nakajima, Yasuto Kondo, Tetsushi Yoshikawa, Atsuo Urisu, Ikuya Tsuge

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background Allergen-specific T-helper type 2 (T H 2) cells play an important role in the development of allergic inflammation; however, investigations of the properties of allergen-specific T cells have been challenging in humans. Despite clear evidence that forkhead box p3 (Foxp3) is expressed in conventional effector T cells, its function has remained unknown. Objective To characterize allergen-specific T H 2 cells in milk allergy, with particular focus on the expression of Foxp3. Methods Twenty-one children with milk allergy and 11 children without milk allergy were studied. Peripheral blood mononuclear cells from subjects were stimulated with milk allergen for 6 hours and analyzed using multicolor flow cytometry to identify CD154 + allergen-specific T-helper cells. Simultaneously, the expression of intracellular cytokines and Foxp3 was analyzed. Results The milk allergy group had significantly larger numbers of milk allergen-specific interleukin (IL)-4- and IL-5-producing CD4 + T cells than the control group. Subjects in the milk allergy group had significantly more CD154 + CD4 + IL-10-producing cells and CD154 + Foxp3 + CD4 + cells than those in the control group. In addition, the number of milk allergen-specific CD154 + Foxp3 + CD4 + cells strongly correlated with that of CD154 + IL4 + CD4 + cells. Bcl-2 expression in CD154 + IL-4 + Foxp3 + T-helper cells was significantly lower compared with that in total CD4 cells. Conclusion Increased numbers of IL-4-producing allergen-specific T-helper cells were found in patients with milk allergy. In addition, Foxp3 was coexpressed with IL-4 in allergen-specific T H 2 cells from patients. This coexpression was associated with lower Bcl-2 levels and could contribute to the phenotype and function of T H 2 cells.

Original languageEnglish
Pages (from-to)503-508
Number of pages6
JournalAnnals of Allergy, Asthma and Immunology
Volume115
Issue number6
DOIs
Publication statusPublished - 01-12-2015

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Th2 Cells
Interleukin-4
Allergens
Milk Hypersensitivity
Milk
Helper-Inducer T-Lymphocytes
T-Lymphocytes
Control Groups
Interleukin-5
Interleukin-10
Blood Cells
Flow Cytometry
Cytokines
Inflammation
Phenotype

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Pulmonary and Respiratory Medicine

Cite this

Yamawaki, Kazuo ; Inuo, Chisato ; Nomura, Takayasu ; Tanaka, Kenichi ; Nakajima, Yoichi ; Kondo, Yasuto ; Yoshikawa, Tetsushi ; Urisu, Atsuo ; Tsuge, Ikuya. / Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3. In: Annals of Allergy, Asthma and Immunology. 2015 ; Vol. 115, No. 6. pp. 503-508.
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title = "Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3",
abstract = "Background Allergen-specific T-helper type 2 (T H 2) cells play an important role in the development of allergic inflammation; however, investigations of the properties of allergen-specific T cells have been challenging in humans. Despite clear evidence that forkhead box p3 (Foxp3) is expressed in conventional effector T cells, its function has remained unknown. Objective To characterize allergen-specific T H 2 cells in milk allergy, with particular focus on the expression of Foxp3. Methods Twenty-one children with milk allergy and 11 children without milk allergy were studied. Peripheral blood mononuclear cells from subjects were stimulated with milk allergen for 6 hours and analyzed using multicolor flow cytometry to identify CD154 + allergen-specific T-helper cells. Simultaneously, the expression of intracellular cytokines and Foxp3 was analyzed. Results The milk allergy group had significantly larger numbers of milk allergen-specific interleukin (IL)-4- and IL-5-producing CD4 + T cells than the control group. Subjects in the milk allergy group had significantly more CD154 + CD4 + IL-10-producing cells and CD154 + Foxp3 + CD4 + cells than those in the control group. In addition, the number of milk allergen-specific CD154 + Foxp3 + CD4 + cells strongly correlated with that of CD154 + IL4 + CD4 + cells. Bcl-2 expression in CD154 + IL-4 + Foxp3 + T-helper cells was significantly lower compared with that in total CD4 cells. Conclusion Increased numbers of IL-4-producing allergen-specific T-helper cells were found in patients with milk allergy. In addition, Foxp3 was coexpressed with IL-4 in allergen-specific T H 2 cells from patients. This coexpression was associated with lower Bcl-2 levels and could contribute to the phenotype and function of T H 2 cells.",
author = "Kazuo Yamawaki and Chisato Inuo and Takayasu Nomura and Kenichi Tanaka and Yoichi Nakajima and Yasuto Kondo and Tetsushi Yoshikawa and Atsuo Urisu and Ikuya Tsuge",
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Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3. / Yamawaki, Kazuo; Inuo, Chisato; Nomura, Takayasu; Tanaka, Kenichi; Nakajima, Yoichi; Kondo, Yasuto; Yoshikawa, Tetsushi; Urisu, Atsuo; Tsuge, Ikuya.

In: Annals of Allergy, Asthma and Immunology, Vol. 115, No. 6, 01.12.2015, p. 503-508.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multicolor flow-cytometric analysis of milk allergen-specific T-helper type 2 cells revealed coexpression of interleukin-4 with Foxp3

AU - Yamawaki, Kazuo

AU - Inuo, Chisato

AU - Nomura, Takayasu

AU - Tanaka, Kenichi

AU - Nakajima, Yoichi

AU - Kondo, Yasuto

AU - Yoshikawa, Tetsushi

AU - Urisu, Atsuo

AU - Tsuge, Ikuya

PY - 2015/12/1

Y1 - 2015/12/1

N2 - Background Allergen-specific T-helper type 2 (T H 2) cells play an important role in the development of allergic inflammation; however, investigations of the properties of allergen-specific T cells have been challenging in humans. Despite clear evidence that forkhead box p3 (Foxp3) is expressed in conventional effector T cells, its function has remained unknown. Objective To characterize allergen-specific T H 2 cells in milk allergy, with particular focus on the expression of Foxp3. Methods Twenty-one children with milk allergy and 11 children without milk allergy were studied. Peripheral blood mononuclear cells from subjects were stimulated with milk allergen for 6 hours and analyzed using multicolor flow cytometry to identify CD154 + allergen-specific T-helper cells. Simultaneously, the expression of intracellular cytokines and Foxp3 was analyzed. Results The milk allergy group had significantly larger numbers of milk allergen-specific interleukin (IL)-4- and IL-5-producing CD4 + T cells than the control group. Subjects in the milk allergy group had significantly more CD154 + CD4 + IL-10-producing cells and CD154 + Foxp3 + CD4 + cells than those in the control group. In addition, the number of milk allergen-specific CD154 + Foxp3 + CD4 + cells strongly correlated with that of CD154 + IL4 + CD4 + cells. Bcl-2 expression in CD154 + IL-4 + Foxp3 + T-helper cells was significantly lower compared with that in total CD4 cells. Conclusion Increased numbers of IL-4-producing allergen-specific T-helper cells were found in patients with milk allergy. In addition, Foxp3 was coexpressed with IL-4 in allergen-specific T H 2 cells from patients. This coexpression was associated with lower Bcl-2 levels and could contribute to the phenotype and function of T H 2 cells.

AB - Background Allergen-specific T-helper type 2 (T H 2) cells play an important role in the development of allergic inflammation; however, investigations of the properties of allergen-specific T cells have been challenging in humans. Despite clear evidence that forkhead box p3 (Foxp3) is expressed in conventional effector T cells, its function has remained unknown. Objective To characterize allergen-specific T H 2 cells in milk allergy, with particular focus on the expression of Foxp3. Methods Twenty-one children with milk allergy and 11 children without milk allergy were studied. Peripheral blood mononuclear cells from subjects were stimulated with milk allergen for 6 hours and analyzed using multicolor flow cytometry to identify CD154 + allergen-specific T-helper cells. Simultaneously, the expression of intracellular cytokines and Foxp3 was analyzed. Results The milk allergy group had significantly larger numbers of milk allergen-specific interleukin (IL)-4- and IL-5-producing CD4 + T cells than the control group. Subjects in the milk allergy group had significantly more CD154 + CD4 + IL-10-producing cells and CD154 + Foxp3 + CD4 + cells than those in the control group. In addition, the number of milk allergen-specific CD154 + Foxp3 + CD4 + cells strongly correlated with that of CD154 + IL4 + CD4 + cells. Bcl-2 expression in CD154 + IL-4 + Foxp3 + T-helper cells was significantly lower compared with that in total CD4 cells. Conclusion Increased numbers of IL-4-producing allergen-specific T-helper cells were found in patients with milk allergy. In addition, Foxp3 was coexpressed with IL-4 in allergen-specific T H 2 cells from patients. This coexpression was associated with lower Bcl-2 levels and could contribute to the phenotype and function of T H 2 cells.

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