Proteomic Profile of Plasma and Mucosal Samples from Patients with Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis

Abstract

Purpose: Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) are life-threatening, immune-complex hypersensitivity adverse drug reactions that affect the skin and mucous membranes, often resulting in significant ocular inflammation and systemic morbidity. The aim of this study was to compare the proteomic and coagulation profiles of sequential mucosal (ocular and oral) and plasma samples from patients with biopsy-confirmed SJS/TENS patients (5) and patients with non-SJS/TENS adverse drug reactions (ADR) (10).

Methods: Multiple plasma samples and swabs from ocular, oral, and skin lesions were obtained from patients with SJS/TEN (n=5). Samples from normal healthy controls were also obtained. The plasma was assayed for thrombin-antithrombin complex (TAT), prothrombin fragment F1.2, plasminogen activator inhibitor-1 (PAI-2), ZYMUPHEN platelet microparticle activity, HEMOCLOT protein C, and STACHROME antithrombin, using ELISA kits as per manufacturer’s instructions. The discharges were isolated following addition of 0.25 ml of saline to each swab and double centrifugation. The discharges and plasma samples were analyzed using SELDI-TOF technique.

Results: Analyses of the SJS/TEN plasma samples revealed a marked increase in the TAT complexes (6.3±5.9µg/ml), F1.2 (430.4±202.4 pmol/L), platelet microparticles (13.1±9.3nM) and protein C levels (90.5±63.4 %), with a corresponding decrease in PAI-1 (53.3±18.8ng/ml) and antithrombin levels (80.7±42.4 %) compared to normal healthy control plasma, suggesting a procoagulant state. Protein chip array of the SJS/TEN skin and oral mucosal samples exhibited two major peaks at 14.2 kDa and 15.6 kDa, in the same molecular weight range as recombinant human granulysin, a molecule implicated in the pathophysiology of SJS/TEN. Additional peaks at 11.7, 13.3 and 14.5 kDa were observed to be upregulated in SJS/TEN patients. These peaks were not present in the normal control group.

Conclusions: Procoagulant factors and unique peaks suggestive of granulysin may lead to the development of targeted therapy aimed to attenuate local and systemic inflammatory processes in patients with SJS/TEN.

Keywords: 663 proteomics