Predominance of type 2 cytokine–producing CD4+ and CD8+ cells in patients with atopic dermatitis,☆☆,,★★

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Abstract

Background: Recently, increased IL-4 and decreased interferon-γ (IFN-γ) production in peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis have been reported by several groups. They measured the total amount of each cytokine in culture supernatants in their studies. These studies suggested a predominance of type 2 cytokine–producing cells in atopic dermatitis. However, it is still unclear whether the cytokine imbalance is the result of an imbalance of specific T-cell subsets or of dysregulation of the cytokine-producing ability in T-cell subsets. Here, we examined frequencies of IL-4–, IFN-γ–, or IL-2–producing CD4+ cells and CD8+ cells in PBMCs from patients with atopic dermatitis at a single cell level by using flow cytometry. Methods: PBMCs from 45 patients with atopic dermatitis and 24 healthy control subjects were stimulated with immobilized anti-CD3 monoclonal antibody for 6 hours in the presence of monensin. Cells were fixed, made permeable, and stained for intracellular cytokines in combination with staining for cell surface markers CD4 and CD8. Results: The frequency of IL-4–producing CD4+ cells and CD8+ cells from patients with atopic dermatitis was significantly higher (p < 0.001) than that from healthy control subjects. In contrast, the frequency of IFN-γ–producing CD4+ cells and CD8+ cells was significantly decreased (p < 0.01) in the patients with atopic dermatitis. The frequency of IL-2–producing cells from patients with atopic dermatitis was comparable to that from healthy control subjects. Conclusion: Our findings indicate that frequencies of type 2 cytokine-producing cells, not only among CD4+ cells but also among CD8+ cells, were significantly higher in patients with atopic dermatitis than in healthy control subjects. (J Allergy Clin Immunol 1997;99:673-82.)

Section snippets

Subjects

Peripheral blood was obtained from 45 patients with AD (19 men and 26 women, aged 25.4 ± 8.6 years [mean ± SD]). AD was diagnosed according to the criteria of Hanifin and Rajka.28 The severity of disease was determined by the criteria of Rajka and Langeland,29 and patients were categorized into three groups. Briefly, symptoms of AD were scored (1 to 3 points in each parameter) by using the following parameters: (1) extent, (2) course (by history), and (3) intensity (disturbance of night's sleep

Frequency of CD4+ and CD8+ cells from patients with AD and from healthy control subjects

To assess the effect of treatment with fixation and permeabilization on staining of surface CD4 and CD8 molecules, we examined frequencies of CD4+ cells and CD8+ cells in treated and untreated samples. Isolated PBMCs were stained with anti-cytokine mAbs, followed by staining of surface CD4 and CD8 antigens with or without fixation in 2% paraformaldehyde and with or without permeabilization in 0.1% saponin. There was no difference in the frequency of CD4+ or CD8+ cells between treated samples

Discussion

In this article, we have demonstrated that the frequency of IL-4–producing CD4+ cells in PBMCs from patients with AD was significantly higher than that from healthy control subjects. In contrast, the frequency of IFN-γ–producing CD4+ cells from patients with AD was significantly lower than that from healthy control subjects when PBMCs were stimulated with immobilized anti-CD3 mAb for 6 hours. Also, among CD8+ cells, the frequency of IL-4–producing cells from patients with AD was significantly

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  • Cited by (0)

    From athe Department of Immunology and Parasitology, bthe Department of Ophthalmology, and cthe Department of Dermatology, School of Medicine, Yokohama City University, Yokohama.

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    Supported in part by Grants-in-Aid from the Japanese Ministry of Education, Science and Culture and by grants from Yokohama City.

    Reprint requests: Mutsuhiko Minami MD, PhD, Department of Immunology and Parasitology, School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawaku, Yokohama, 236 Japan.

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