Ann Dermatol.  2014 Jun;26(3):308-313.

Comparative Analysis of Human Epidermal and Peripheral Blood gammadelta T Cell Cytokine Profiles

• Kim K
• Han J
• Lee TR
• Shin DW
• Chang H
• Cho AR
• Choi SJ
• Jo SJ
• Kwon O

Abstract

BACKGROUND
Human epidermal gammadelta T cells are known to play crucial roles in the defense and homeostasis of the skin. However, their precise mechanism of action in skin inflammation remains less clear.
OBJECTIVE
In this study, we analyzed the cytokine expression profile of human epidermal gammadelta T cells and compared it to that of peripheral blood gammadelta T cells to investigate the specific activity of epidermal gammadelta T cells in modulating skin inflammation.
METHODS
We isolated gammadelta T cells from epidermal tissue or peripheral blood obtained from healthy volunteers. Isolated gammadelta T cells were stimulated using immobilized anti-CD3 antibody and interleukin-2 plus phytohaemagglutinin, and were then analyzed using a cytokine array kit.
RESULTS
Both epidermal and peripheral blood gammadelta T cells produced comparable levels of granulocyte-macrophage colony-stimulating factor, I-309, interferon-gamma, macrophage migration inhibitory factor, macrophage inflammatory protein-1alpha, and chemokine (C-C) ligand 5. The epidermal gammadelta T cells produced significantly higher levels of interleukin-4, -8, -13, and macrophage inflammatory protein-1beta than the peripheral blood gammadelta T cells did. Notably, the epidermal gammadelta T cells produced several hundred-fold higher levels of interleukin-13 than interleukin-4.
CONCLUSION
These results suggest that the epidermal gammadelta T cells have a stronger potential to participate in the Th2-type response than the peripheral blood gammadelta T cells do. Furthermore, epidermal gammadelta T cells might play an important role in the pathogenesis of Th2-dominant skin diseases because of their active production of interleukin-13.

Keyword

Epidermis; Interleukin-4; Interleukin-13; gammadelta T cells

MeSH Terms

Epidermis
Granulocyte-Macrophage Colony-Stimulating Factor
Healthy Volunteers
Homeostasis
Humans
Inflammation
Interferon-gamma
Interleukin-13
Interleukin-2
Interleukin-4
Macrophages
Skin
Skin Diseases
T-Lymphocytes
Granulocyte-Macrophage Colony-Stimulating Factor
Interferon-gamma
Interleukin-13
Interleukin-2
Interleukin-4

Figure

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  Fig. 1 Flow cytometric analysis of the purified epidermal and peripheral blood γδ T cells. Purified γδ T cells obtained from human epidermal tissue (A) and from the peripheral blood (B) of healthy volunteers were stained using a fluorescein-conjugated anti-CD3 monoclonal antibody (mAb) and APC-conjugated anti-γδ T cell receptor mAb or the corresponding fluorescently conjugated isotype-matched control Abs. Three independent experiments were performed for epidermal cells and peripheral blood cells respectively, and the representative results are presented. FITC: fluorescein isothiocyanate.

• 

  Fig. 2 Comparison of cytokine expression profiles of epidermal and peripheral blood γδ T cells. Epidermal γδ T cells were isolated from the epidermes of three healthy volunteers (A~C) by using the method described in the Materials and Methods section. Peripheral blood γδ T cells were isolated from the peripheral blood of three healthy volunteers (D~F) by using conventional Histopaque 1077 gradient centrifugation, followed by magnetic-activated cell sorting. Epidermal and peripheral blood γδ T cells were stimulated using immobilized anti-CD3 antibody and IL-2 plus phytohaemagglutinin. The culture supernatants were analyzed after 36 hours by using the Human Cytokine Array Panel A. (A~F) cytokine array membranes. (G, H) The blot intensities on the array membranes were measured using a gel documentation system. Data are presented as the mean±standard deviation. Statistical analyses were performed using the Student's t-test. C5a: complement component 5a, G-CSF: granulocyte colony-stimulating factor, GM-CSF: granulocyte-macrophage colony stimulating factor, GROα: growth-related oncogene α, sICAM-1: soluble intercellular adhesion molecule-1, IFNγ: interferon-γ, IL: interleukin, IP-10: interferon gamma-induced protein 10, I-TAC: interferon-inducible T cell α chemoattractant, MCP-1: monocyte chemotactic protein 1, MIP: macrophage inflammatory protein, RANTES: regulated on activation, normal T cell expressed and secreted, SDF-1: stromal cell-derived factor 1, TNF-α: tumor necrosis factor α, sTREM-1: soluble triggering receptor expressed on myeloid cells 1. *p<0.05, epidermal vs. peripheral blood γδ T cells.

• 

  Fig. 3 Quantitative analyses of interleukin-4 and interleukin-13 production in epidermal γδ T cells and peripheral blood γδ T cells. Epidermal and peripheral blood γδ T cells were each isolated from three healthy volunteers. Cells were stimulated using immobilized anti-CD3 antibody and interleukin-2 plusphytohaemagglutinin, or cultured without any stimulation. The culture supernatants were analyzed after 36 hours by using ELISAs. Data are presented as the mean±standard deviation of three independent experiments. Statistical analyses were performed using the Student's t-test. EP: epidermal, PB: peripheral blood. *p<0.05, epidermal vs. peripheral γδ T cells.

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