Abstract
BackgroundPaediatric systemic lupus erythematosus (pSLE) exhibits an aggressive clinical phenotype with severe complications and overall poor prognosis. The aim of this study was to analyse differential expression of low molecular weight (LMW) serum protein molecules of pSLE patients with active disease in comparison to sera of healthy age matched controls. Further, some of the differential expressed spots were characterised and identified by Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) and liquid chromatography (LC-MS).Methods2D-PAGE was performed using pooled sera of active pSLE and age matched healthy controls. Gels were silver-stained and differentially expressed protein spots were detected by automated image master platinum 2D software. 79 ± 17 protein spots were detected for control gels and 78 ± 17 protein spots for patient gels. Of these eleven protein spots were selected randomly and characterized by MALDI-TOF MS (five protein spots) and LC MS (six protein spots) techniques.ResultsOut of the 11 protein spots, 5 protein spots were significantly upregulated viz., leiomodin 2 (LMOD2); epidermal cytokeratin 2; immunoglobulin kappa light chain variable region; keratin 1 and transthyretin (TTR). Three protein spots were significantly down regulated e.g., apolipoprotein A1 (APOA1); chain B human complement component C3c; campath antibody antigen complex. Two protein spots (complement component C3; retinol binding protein (RBP) were found to be expressed only in disease and one protein spot cyclohydrolase 2 was only expressed in controls.ConclusionsWe conclude that 2-D maps of patients with active pSLE and controls differ significantly. In this pilot study, using proteomic approach we have identified differential expressed proteins (of LMW) e.g., RBP, LMOD 2, TTR, Component C3c Chain B and APO A1. However, in future, further studies need to confirm the physiological and pathological role of these proteins in similar cohorts of pSLE.
Highlights
Paediatric systemic lupus erythematosus exhibits an aggressive clinical phenotype with severe complications and overall poor prognosis
The diagnosis of Paediatric systemic lupus erythematosus (pSLE) patients was based on the American College of Rheumatology (ACR) criteria for classification [15]
We have described the increased expression of Retinol Binding Protein (RBP) protein spots in pooled sera of pSLE patients with active disease and compared them with controls
Summary
Paediatric systemic lupus erythematosus (pSLE) exhibits an aggressive clinical phenotype with severe complications and overall poor prognosis. A proteomic approach is necessary to find proteins expressed in the disease states, which may serve as diagnostic or prognostic marker, or serve as targets for therapeutics. Such an approach has yielded newer diagnostic proteins in the other autoimmune diseases viz., myelin protein Po in autoimmune inner ear disease [1], heterogeneous nuclear ribonucleoprotein A2/B1 in autoimmune hepatitis, ?-enolase in Behcet’s disease [2], ?-enolase and its citrullinated molecule in rheumatoid arthritis [3,4,5,6,7]. Protein function can be regulated by posttranslational modifications by enzymes such as kinases or proteases
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