Ann Dermatol.  2014 Oct;26(5):576-583. 10.5021/ad.2014.26.5.576.

Preferential Recognition of Hydroxyl Radical-Modified Superoxide Dismutase by Circulating Autoantibodies in Patients with Alopecia Areata

• Alzolibani AA

• KMID: 2265490
• DOI: http://doi.org/10.5021/ad.2014.26.5.576

Abstract

BACKGROUND
Alopecia areata (AA) is a common form of localized, non-scarring hair loss. The cause of AA is unknown but reports suggest an autoimmune etiology, where oxygen free radicals play an important role.
OBJECTIVE
The aim of this study was to investigate the role of a hydroxyl radicals (.OH)-modified antioxidant enzyme, superoxide dismutase (SOD), in AA autoimmunity.
METHODS
SOD was modified by .OH radicals. Binding characteristics of autoantibodies in AA patients (n=26) against .OH-modified SOD (.OH-SOD) were evaluated by immunoassays and the results were compared with those of healthy, age-matched controls (n=30). The effects of .OH radicals on immunoglobulin G (IgG) isolated from AA patients were studied.
RESULTS
Highly specific binding to .OH-SOD was observed in 32% of the samples of patient sera, whereas normal human sera showed negligible binding with either antigen. Competitive inhibition immunoassays reiterated the results from direct binding. Protein-A-purified IgG from AA patients (AA-IgG) also showed strong binding to .OH-SOD as compared to IgG from normal human controls (p<0.001). In addition, AA-IgG from patients with alopecia universalis recognized .OH-SOD to a greater extent than did AA-IgG from patients with the patchy, persistent type of alopecia. Furthermore, sera from AA patients had lower levels of SOD activity as compared to control sera.
CONCLUSION
This is the first report showing an association between .OH-modified SOD and AA. These novel results demonstrate that .OH radical-mediated changes in SOD present unique neo-epitopes that might contribute to antigen-driven antibody induction in AA.

Keyword

Alopecia areata; Autoimmunity; .OH-SOD; Reactive oxygen species; Superoxide dismutase

MeSH Terms

Alopecia
Alopecia Areata*
Autoantibodies*
Autoimmunity
Free Radicals
Hair
Humans
Immunoassay
Immunoglobulin G
Oxygen
Reactive Oxygen Species
Superoxide Dismutase*
Autoantibodies
Free Radicals
Immunoglobulin G
Oxygen
Reactive Oxygen Species
Superoxide Dismutase

Figure

• 

  Fig. 1 Direct binding of alopecia areata antibodies to reactive oxygen species-modified superoxide dismutase (ROS-SOD). (A) Levels of circulating antibodies in alopecia areata (AA) patients, binding to ROS-SOD and native SOD (nSOD). Anti-ROS-SOD antibodies versus anti-nSOD antibodies, p<0.01 in AA patients. (B) Levels of circulating antibodies in normal human (NH) subjects binding to ROS-SOD and nSOD. Anti-ROS-SOD antibodies versus anti-nSOD antibodies, p>0.05 in NH subjects. Microtiter plates were individually coated with ROS-SOD (10 µg/ml) and nSOD (10 µg/ml). OD: optical density or absorbance.

• 

  Fig. 2 Affinity-purification of immunoglobulin G (IgG). Elution profile of IgG on a Protein A agarose affinity column.

• 

  Fig. 3 Binding of Protein A-purified alopecia areata (AA) immunoglobulin G (IgG) to reactive oxygen species-modified superoxide dismutase (ROS-SOD) and native superoxide dismutase. (A) Binding characteristic of AA-IgG and normal human (NH)-IgG to ROS-SOD. (B) Binding characteristics of AA-IgG and NH-IgG to native SOD (nSOD). IgG from 12 AA patients and 12 NH subjects were analyzed by direct-binding enzyme-linked immunosorbent assay. Microtiter plates were individually coated with ROS-SOD (10 µg/ml) and nSOD (10 µg/ml). For ROS-SOD: AA patients vs. #p<0.01; For nSOD: AA patients vs. NH subjects, p>0.05. OD: optical density or absorbance.

• 

  Fig. 4 Disease-related decrease in superoxide dismutase (SOD) activity in patients with alopecia areata (AA). Serum levels of SOD activity in AA patients (n=25) compared with normal human (NH) subjects (n=26). Each histogram represents the mean±standard error of the mean. SOD activity in AA patients vs. SOD activity in NH subjects, p<0.05.

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