J Korean Med Sci.  2006 Apr;21(2):315-323. 10.3346/jkms.2006.21.2.315.

Role of Staphylococcal Superantigen in Atopic Dermatitis: Influence on Keratinocytes

• Kim KH
• Han JH
• Chung JH
• Cho KH
• Eun HC

• KMID: 1781842
• DOI: http://doi.org/10.3346/jkms.2006.21.2.315

Abstract

Staphylococcus aureus may perform an crucial function in atopic dermatitis (AD), via the secretion of superantigens, including staphylococcal enterotoxins (SE) A or B, and toxic shock syndrome toxin-1 (TSST-1). Dysregulated cytokine production by keratinocytes (KCs) upon exposure to staphylococcal superantigens (SsAgs) may be principally involved in the pathophysiology of AD. We hypothesized that lesional KCs from AD may react differently to SsAgs compared to nonlesional skin or normal skin from nonatopics. We conducted a comparison of HLA-DR or CD1a expression in lesional skin as opposed to that in nonlesional or normal skin by immunohistochemistry (IHC). We also compared, using ELISA, the levels of IL-1alpha, IL-1beta, and TNF-alpha secreted by cultured KCs from lesional, nonlesional, and normal skin, after the addition of SEA, SEB and TSST-1. IHC revealed that both HLA-DR and CD1a expression increased significantly in the epidermis of lesional skin versus nonlesional or normal skin in quite a similar manner. IL-1alpha, IL-1beta, and TNF-alpha secretion was also significantly elevated in the cultured KCs from lesional skin after the addition of SsAgs. Our results indicated that KCs from lesional skin appear to react differently to SsAgs and increased proinflammatory cytokine production in response to SsAgs may contribute to the pathogenesis of AD.

Keyword

Dermatitis, Atopic; Staphylococcal Superantigens; Staphylococcal enterotoxin B; HLA-DR Antigens; Interleukin-1; Tissue Necrosis Factor-alpha

MeSH Terms

Tumor Necrosis Factor-alpha/biosynthesis/genetics
*Superantigens/administration & dosage/immunology
Staphylococcus aureus/*immunology/pathogenicity
Male
Keratinocytes/immunology/*microbiology
Interleukin-1/biosynthesis/genetics
Inflammation Mediators/metabolism
Humans
HLA-DR Antigens/metabolism
Enterotoxins/administration & dosage/immunology
Dermatitis, Atopic/etiology/immunology/*microbiology
DNA, Complementary/genetics
Case-Control Studies
Base Sequence
Bacterial Toxins/administration & dosage/immunology
Antigens, CD1/metabolism
Adult

Figure

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  Fig. 1 (A) Numbers of HLA-DR (+) cells in the epidermis increased significantly in lesional skin, as compared to those from nonlesional skin from AD patients and from normal skin from nonatopic controls (p<0.05). (B) Numbers of CD1a cells in the epidermis also increased significantly quite similarly to those of HLA-DR (+) cells in terms of their increased numbers and their locations in lesional skin, as compared to those from nonlesional skin from AD patients and from normal skin from nonatopic controls (p<0.05).

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  Fig. 2 No significant cellular cytotoxicity was detected with any of the tested SsAgs concentrations.

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  Fig. 3 The levels of IL-1α released from cultured KCs in response to SsAgs. Only 1.0 µg/mL of SEA and SEB, and 0.1 and 1.0 µg/mL of TSST-1 significantly increased IL-1α release from cultured atopic KCs vs. the atopic controls or normal KCs (p<0.05). In the case of TSST-1, 1.0 µg/mL significantly increased IL-1α release from atopic KCs vs. 0.1 µg/mL (p<0.05). (A) IL-1α release in response to SEA, (B) IL-1α release in response to SEB, (C) IL-1α release in response to TSST-1. *comparison among skin samples. †comparison among doses of toxins.

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  Fig. 4 IL-1β levels released from cultured KCs and IL-1β mRNA in response to SsAgs. Both concentrations (0.1 and 1.0 µg/mL) of SEA, SEB, and TSST-1 resulted in significant increases in IL-1β release from cultured atopic KCs and atopic control KCs vs. the negative control KCs (p<0.05). The increased secretion of IL-1β from each of the KCs proceeded in a clearly dose-dependent manner. The most pronounced changes were observed with TSST-1. Increased IL-1β mRNA in the atopic KCs and atopic control KCs vs. the negative control KCs was also observed, particularly in response to SEB and TSST-1 stimulation. (A) IL-1β release in response to SEA, (B) IL-1β release in response to SEB, (C) IL-1β release in response to TSST-1, (D) IL-1β mRNA levels measured by semi-quantitative RT-PCR. *comparison among skin samples, †comparison among doses of toxins.

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  Fig. 5 TNF-α release levels from the cultured KCs and TNF-α mRNA in response to SsAgs. Both tested concentrations (0.1 and 1.0 µg/mL) of SEA, SEB, and TSST-1 resulted in significant increases in the release of TNF-α from cultured atopic KCs and atopic control KCs vs. the negative control KCs (p<0.05). Increased TNF-α mRNA in the atopic KCs vs. the atopic control KCs and negative control KCs was observed, particularly in the cases of 1.0 µg/mLof SEA and SEB stimulations. (A) TNF-α release in response to SEA, (B) TNF-α release in response to SEB, (C) TNF-α release in response to TSST-1, (D) TNF-α mRNA levels measured by semi-quantitative RT-PCR. *comparison among skin samples, †comparison among doses of toxins.

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