Abstract
Antigen-presenting cells (APC) are thought to play an important role in the pathogenesis of drug-induced immune reactions. Various pathological factors can activate APC and therefore influence the immune equilibrium. It is interesting that several diseases have been associated with an increased rate of drug allergy. The aim of this project was to evaluate the impact of such “danger signals” on sulfamethoxazole (SMX) metabolism in human APC (peripheral blood mononuclear cells, Epstein-Barr virus-modified B lymphocytes, monocyte-derived dendritic cells, and two cell lines). APC were incubated with SMX (100 μM–2 mM; 5 min–24 h), in the presence of pathological factors: bacterial endotoxins (lipopolysaccharide and staphylococcal enterotoxin B), flu viral proteins, cytokines [interleukin (IL)-1β, IL-6, IL-10; tumor necrosis factor-α; interferon-γ; and transforming growth factor-β], inflammatory molecules (prostaglandin E2, human serum complement, and activated protein C), oxidants (buthionine sulfoximine and H2O2), and hyperthermia (37.5–39.5°C). Adduct formation was evaluated by enzyme-linked immunosorbent assay and confocal microscopy. SMX-protein adduct formation was time- and concentration-dependent for each cell type tested, in both physiological and danger conditions. A danger environment significantly increased the formation of SMX-protein adducts and significantly shortened the delay for their detection. An additive effect was observed with a combination of danger signals. Dimedone (chemical selectively binding cysteine sulfenic acid) and antioxidants decreased both baseline and danger-enhanced SMX-adduct formation. Various enzyme inhibitors were associated with a significant decrease in SMX-adduct levels, with a pattern varying depending on the cell type and the culture conditions. These results illustrate that danger signals enhance the formation of intracellular SMX-protein adducts in human APC. These findings might be relevant to the increased frequency of drug allergy in certain disease states.
- APC, antigen-presenting cells
- SMX, sulfamethoxazole
- HIV, human immunodeficiency virus
- SMX-HA, sulfamethoxazole-hydroxylamine
- SMX-NO, sulfamethoxazole-nitroso
- Mo-DC, monocyte-derived dendritic cells
- LPS, lipopolysaccharide
- PMA, phorbol 12-myristate 13-acetate
- PBMC, peripheral blood mononuclear cells
- IL, interleukin
- EBV, Epstein-Barr virus
- ELISA, enzyme-linked immunosorbent assay
- SEB, staphylococcal enterotoxin B
- TNF, tumor necrosis factor
- TGF, transforming growth factor
- IFN, interferon
- PGE, prostaglandin E
- BSO, buthionine sulfoximine
- GSH, glutathione
- ABT, aminobenzotriazole
- MTZ, methimazole
- COX, cyclooxygenase
- ABH, aminobenzoic hydrazide
- MPO, myeloperoxidase
- OD, optical density
- AA, ascorbic acid
- DMSO, dimethyl sulfoxide.
Footnotes
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This work was supported by the Wellcome Trust [Grant 078598/Z/05/Z] (as part of the Centre for Drug Safety Science supported by the Medical Research Council [Grant G0700654]).
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.109.155374
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↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
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ABBREVIATIONS:
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↵1 Current affiliation: Veterinary College of Illinois, Urbana-Champaign, Illinois.
- Received April 20, 2009.
- Accepted August 6, 2009.
- © 2009 by The American Society for Pharmacology and Experimental Therapeutics
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