June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Proteomic Analysis of Plasma and Mucosal Samples from Patients with Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis
Author Affiliations & Notes
  • Julia Malalis
    Ophthalmology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Christine Mata
    Ophthalmology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Daniel Kahn
    Pathology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Amy Lin
    Ophthalmology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Michael Mosier
    Surgery, Loyola University Medical Center, Maywood, IL
  • Charles Bouchard
    Ophthalmology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Josephine Cunanan
    Pathology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Omer Iqbal
    Ophthalmology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
    Pathology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Debra Hoppensteadt
    Pathology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Jawed Fareed
    Pathology, Loyola University Chicago Stritch School of Medicine, Maywood, IL
  • Footnotes
    Commercial Relationships Julia Malalis, None; Christine Mata, None; Daniel Kahn, None; Amy Lin, None; Michael Mosier, None; Charles Bouchard, None; Josephine Cunanan, None; Omer Iqbal, None; Debra Hoppensteadt, None; Jawed Fareed, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2062. doi:
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      Julia Malalis, Christine Mata, Daniel Kahn, Amy Lin, Michael Mosier, Charles Bouchard, Josephine Cunanan, Omer Iqbal, Debra Hoppensteadt, Jawed Fareed; Proteomic Analysis of Plasma and Mucosal Samples from Patients with Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2062.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) are life-threatening, immune-complex hypersensitivity reactions that affect the skin and mucous membranes, often resulting in significant ocular inflammation. The purpose of this study was to determine and compare the proteomic and coagulation profile of plasma and mucosal discharge samples in affected and unaffected patients.

Methods: Following institutional review board approval, plasma and swabs from ocular, oral, and skin lesions were obtained from patients with clinically suspected SJS/TEN (n=5). Two patients had biopsy-proven SJS/TEN. Three patients had alternative skin diagnoses not consistent with SJS/TEN and were considered abnormal controls. Samples from normal healthy controls were also obtained. Following centrifugation, the plasma was frozen at -70°C and later thawed and analyzed to determine thrombin-antithrombin complex (TAT, Dade®, Marburg, Germany), fibrinopeptide (F1.2, Dade®), plasminogen activator inhibitor-1 (PAI-1, Diagnostica Stago®, Asnieres Sur Seine, France), ZYMUPHEN platelet microparticle activity (Hyphen® BioMed, Neuville-Sur Oise, France), HEMOCLOT protein C (Stago®), and STACHROME antithrombin (Stago®), using ELISA kits as per manufacturer’s instructions. The swabs were immediately frozen at -70°C and later thawed. 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 control human 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. These peaks were not present in the 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: 490 cytokines/chemokines • 557 inflammation • 480 cornea: basic science  
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