J Bacteriol Virol.  2017 Sep;47(3):132-138. 10.4167/jbv.2017.47.3.132.

The Effects of Staphylococci on the Degranulation of Human Mast Cell-1

• Jang IT
• Yang M
• Jo EK
• Kim HJ
• Park JK

• KMID: 2392961
• DOI: http://doi.org/10.4167/jbv.2017.47.3.132

Abstract

Atopic dermatitis (AD) is characterized by disturbances in epidermal barrier functions and the hyperactive immune response. Staphylococcus aureus (S. aureus) can be cultured from 90% of AD skin lesions and can exacerbate or contribute to the persistent skin inflammation in AD by secreting toxins with superantigenic properties. Superantigens can induce mast cell (MC) degranulation after penetrating the epidermal barrier. The role of MCs in AD is suggested by the increase in the MC number and MC activation. MCs are activated for degranulation and mediator release by allergens that cross-link IgE molecules or by microbial products. Therefore, MCs may be critically involved in the pathogenesis of AD. However, the understanding mechanisms of MC degranulation by S. aureus in relation to AD have still not been fully elucidated. In this study, we found that live S. aureus or methicillin-resistant S. aureus (MRSA) but not heat-killed bacteria induced MC degranulation. The heat-treatment partially inhibited MC degranulation by conditioned media (CM) of S. aureus or MRSA. The calcium chelator ethylene glycol tetraacetic acid (EGTA) did not block MC degranulation induced by live S. aureus or MRSA, but EGTA-treatment partially inhibited MC degranulation by CM from S. aureus or MRSA. These results suggest that live S. aureus and MRSA can degranulate MCs via direct interaction which may be important role in AD.

Keyword

Staphylococci; Mast cell; β-hexosaminidase; Atopic dermatitis

MeSH Terms

Allergens
Bacteria
Calcium
Culture Media, Conditioned
Dermatitis, Atopic
Egtazic Acid
Humans*
Immunoglobulin E
Inflammation
Mast Cells
Methicillin Resistance
Methicillin-Resistant Staphylococcus aureus
Skin
Staphylococcus aureus
Superantigens
Allergens
Calcium
Culture Media, Conditioned
Egtazic Acid
Immunoglobulin E
Superantigens

Figure

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  Figure 1. MRSA induces HMC-1 degranulation. β-hexosa-minidase activity (%) was determined from HMC-1 cultured with MRSA (MOI = 100) for indicated time (minutes). Data represent means ± s.d. of triplicate independent experiments.

• 

  Figure 2. The conditioned media from MRSA induces HMC-1 degranulation. β-hexosaminidase activity (%) was determined from HMC-1 stimulated with different concentrations of conditioned media of MRSA. Data represent means ± s.d. of triplicate independent experiments. ∗⦤0.05.

• 

  Figure 3. Differences in HMC-1 degranulations between live and heat-killed Staphylococci. β-hexosaminidase activity (%) was determined from HMC-1 stimulated with heat-killed S. epidermidis (SE), S. aureus (SA) or MRSA, and heat-killed bacteria-induced β-hexosaminidase activity (%) was compared with that of live SE, SA or MRSA. Data represent means ± s.d. of triplicate independent experiments. ∗∗∗⦤0.001. ns, no significance.

• 

  Figure 4. Differences in HMC-1 degranulations between conditioned media and heat-inactivated conditioned media from Staphylococci. β-hexosaminidase activity (%) was determined from HMC-1 stimulated with conditioned media (CM) from S. epidermidis (SE), S. aureus (SA) or MRSA, and conditioned media-induced β-hexosaminidase activity (%) was compared with that of heat-inactivated conditioned media (CM). Data represent means ± s.d. of triplicate independent experiments. ∗⦤0.05, ∗∗⦤0.01, ∗∗∗⦤0.001. ns, no significance.

• 

  Figure 5. Differences in HMC-1 degranulations between EGTA pretreated HMC-1 and non-EGTA pretreated HMC-1 induced by Staphylococci. HMC-1 was either pretreated with EGTA or not, and β-hexosaminidase activity (%) induced by S. epidermidis (SE), S. aureus (SA) or MRSA was determined. Data represent means ± s.d. of triplicate independent experiments. ∗∗⦤0.01. ns, no significance.

• 

  Figure 6. Differences in HMC-1 degranulations between EGTA pretreated HMC-1 and non-EGTA pretreated HMC-1 induced by conditioned media (CM) from Staphylococci. HMC-1 was either pretreated with EGTA or not, and β-hexosaminidase activity (%) induced by conditioned media (CM) from S. epidermidis (SE), S. aureus (SA) or MRSA was determined. Data represent means ± s.d. of triplicate independent experiments. ∗∗⦤0.01. ns, no significance.

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