Mechanisms of allergyInduction of corticosteroid insensitivity in human PBMCs by microbial superantigens☆,☆☆
Section snippets
Lymphocyte proliferation assays
PBMCs from 7 healthy donors were isolated from heparinized blood by Ficoll-Hypaque (Pharmacia Biotech, Piscataway, NJ) gradient centrifugation and suspended in RPMI 1640 media (Bio-Whittaker, Walkersville, Md) containing 10% heat-inactivated (30 minutes at 56°C) FCS, 40 μmol/L L -glutamine, 100 U/mL penicillin G sodium, 100 U/mL streptomycin, and 20 mmol/L HEPES buffer solution (GIBCO BRL Life Technologies, Grand Island, NY). PBMCs were plated at 2 × 105 cells per well in 96-well round-bottom
Effect of DEX on in vitro incubation of PBMCs with SAgs
Because SAgs have been described to contribute to the pathogenesis of various chronic inflammatory diseases frequently associated with a reduced response to GC therapy, we assessed the GC sensitivity of normal PBMCs stimulated with microbial SAgs. PBMCs from 7 healthy volunteers were stimulated with the prototypic SAgs SEB, TSST-1, and SEE compared with PHA in the presence of increasing concentrations of DEX. In Fig 1 the responses are expressed as percent of the corresponding culture with the
GRβ expression after stimulation with SEB
Because high-level GRβ expression has been associated with GC insensitivity,16 it was of interest to determine whether SAgs induced GRβ. In these experiments, PBMCs from healthy control subjects were evaluated for GRβ expression by immunocytochemistry after stimulation with SEB compared with PHA. Positive staining for GRβ was observed on all samples stained with anti-GRβ (Fig 3, A and B ) but not the Ig isotype control (Fig 3, C ).
DISCUSSION
Understanding the mechanism by which SAgs might contribute to poor disease control in SAg-triggered chronic inflammatory diseases would be of great interest. In this study we therefore investigated whether SAgs could induce GC insensitivity because GCs play a crucial role in the control of chronic inflammation. Our data demonstrate that prototypic SAgs can render PBMCs of healthy individuals highly resistant to the T-cell inhibitory effect of DEX. PBMCs of the same individuals, incubated with
Acknowledgements
We thank Maureen Sandoval for her excellent support in preparing this manuscript.
References (26)
- et al.
Superantigens and their potential role in human disease
Adv Immunol
(1993) - et al.
Increased T-cell receptor vβ8+ T cells in bronchoalveolar lavage fluid of subjects with poorly controlled asthma: a potential role for microbial superantigens
J Allergy Clin Immunol
(1999) - et al.
Evidence for superantigen involvement in skin homing of T cells in atopic dermatitis
J Invest Dermatol
(1999) - et al.
Staphylococcal toxins and protein A differentially induce cytotoxicity and release of tumor necrosis factor-alpha from human keratinocytes
J Invest Dermatol
(1996) - et al.
Psoriasis: a T-cell–mediated autoimmune disease induced by streptococcal superantigens?
Immunol Today
(1995) - et al.
Effects of glucocorticoids on lymphocyte activation in patients with steroid-sensitive and steroid-resistant asthma
J Allergy Clin Immunol
(1996) - et al.
The human glucocorticoid receptor beta isoform: expression, biochemical properties, and putative function
J Biol Chem
(1996) - et al.
Human eosinophils synthesize and secrete interleukin-6, in vitro
Blood
(1992) - et al.
The dominant negative activity of the human glucocorticoid receptor beta isoform: specificity and mechanisms of action
J Biol Chem
(1999) - et al.
An open-label study of high-dose intravenous immunoglobulin in severe childhood asthma
J Allergy Clin Immunol
(1991)
Selective expansion of T cells expressing V beta 2 in toxic shock syndrome
J Exp Med
Characterization of T cell repertoire changes in acute Kawasaki disease
J Exp Med
Evidence for the effects of a superantigen in rheumatoid arthritis
Science
Cited by (202)
Atopic Dermatitis and Allergic Contact Dermatitis
2022, Allergy EssentialsGlucocorticoid insensitivity by staphylococcal enterotoxin B in keratinocytes of allergic dermatitis is associated with impaired nuclear translocation of the Glucocorticoid Receptor α
2018, Journal of Dermatological ScienceCitation Excerpt :S. aureus on the AD skin can secrete various exotoxins (SE), including SEA, SEB, SEC, SED, and TSST-1, these compounds are also referred to as staphylococcal superantigens (SsAgs) [4]. Several studies have suggested that superantigens are associated with impaired in vitro responsiveness of human PBMCs [5], T cells [6] and monocytes [7] to the suppressive effects of GC. Moreover, the degree of colonization has been found to be associated with disease severity and has also been implicated in the development of insensitivity to glucocorticoids [8–10].
Staphylococcus aureus colonization is associated with increased inhaled corticosteroid requirements in patients with atopic dermatitis and asthma
2017, Journal of Allergy and Clinical Immunology: In PracticeWhen does atopic dermatitis warrant systemic therapy? Recommendations from an expert panel of the International Eczema Council
2017, Journal of the American Academy of Dermatology
- ☆
Supported in part by National Institutes of Health Public Health Services research grants No. HL36577, AR 41256, and HL37260; General Clinical Research Center grant No. 5 MO1 RR00051 from the Division of Research Resources, University of Colorado Cancer Center; an American Lung Association Asthma Research Center grant; the Medical Research Council of Canada; and Inspiraplex. P. J. H. is the recipient of the 1997 AAAAI-Glaxo Respiratory Diseases Research Award.
- ☆☆
Reprint requests: Donald Y. M. Leung, MD, PhD, National Jewish Medical and Research Center, Department of Pediatrics, Room K926, 1400 Jackson St, Denver, CO 80206.