Proteomic Analysis Of PBMC From Patients With Systemic Lupus Erythematosus

Objective: Systemic lupus erythematosus(SLE) is the typical autoimmune disease, of which immunopathogenesis and definition were still in a mess due to complicated clinical symptoms and multiple systems involved. The diagnosis criteria ,depending on the clinical symptoms already exist, rather than autoimmunity state, is not so perfect: when the clinical symptom is mild and untypical, it's easy to be ignored; while it gets worse and easy to recognize, there are always some systemic symptoms like autoantibody, hypocomplementemia, nephritis or encephalopathy etc, even becoming decompensation and making very poor prognosis. Therefore, early diagnosis is very important. In these decades, proteomics is a new born baby of molecular biology, who brought some brand-new methods. During progress, proteomics unveils to make a good contribution and a shiny future on pathomechanism, early diagnosis, drug target research of SLE. Update, the majority of research are focused on sera and urine, which easily obtained. Respectively proteome discovery about immunological cells is barely seen. Considering the wild involvements of pathogenesis, we choose peripheral blood monouclear cells( PBMC), some polyclones including T cells, B cells, NK cells, etc, as our objects. We adopt two-dimensional electrophoresis(2-DE) and Ion Trap high performance liquid chromatography chip tandem mass spectrometer(Ion Trap HPLC-Chip-MS/MS) to discover the comparative proteome of PBMC, in order to chase some useful biomarks.Methods: PBMC protein samples were extracted from 14 SLE patients and 9 healthy controls. The protein was separated by immobilized pH gradient two-dimensional gel electrophoresis. The 2-D electrophoretogram was analyzed using PDquest7.1 software. The differential spots were identified by Ion Trap HPLC-Chip-MS/MS and database searching was performed on the internet.Results:1. The 2-D electrophoretograms were established stably and reproducibly. 2. Match gel eletrophoretogram showed 1084 match spots in 1 SLE sample and 4-in-1 healthy control sample, of which 71 and 17 spots were overexpressed or unique in SLE group , while in control, these data were replaced by 45 and 20 respectively.3. We compared these macth spots one by one and made the reproducible 30 ones selected and through the progress of Ion Trap HPLC-Chip-MS/MS. 28 of them were identified and represented 27 kinds of protein. They were glyceraldehydes-3-phosphate dehydrogenase,tyrosyl-tRNA synthetase, glycogen phosphorylase, transketolase, pyruvate kinase isozymes M1/M2, triosephosphate isomerase, elongation factor 1-alpha 1, phosphoglyceric kinase 1,α-enolase, zyxin, gelsolin precursor, praline-serine-threonine phosphatase-interacting protein 2, adenyl cyclase-associated protein 1, filamin C, filamin A, WD repeat-containing protein 1, talin 1, tubulin beta chain, integrin-linked protein kinase, PDZ and LIM domain protein 7, tyrosine protein kinase CSK, osteoclast-stimulating factor 1, annexin A5, T-complex protein 1 subunit zeta, heat shock protein beta 1, heat shock cognate 71kDa protein, and dynamin-1 like protein. The expressions of 8 kinds in the 27 ones, which included gelsolin precursor, phosphoglyceric kinase 1, WD repeat-containing protein 1,α-enolase, osteoclast-stimulating factor 1, tubulin beta chain, annexin A5, heat shock cognate 71kDa protein, were aggressive in patients suffered from SLE ,others rather inhibited.Conclusion:1. We apply proteomics techniques in screening and identifying some protein related to PBMC of SLE and chase 27 kinds of protein(triosephosphate isomerase, annexin A5, heat shock cognate 71kDa protein and so on) with expression variation, which might be about to become several significant biomarks.2. There maybe some specific clusters of biomarks which represent different clusters of clinical symptoms. Certain protein might be just observed on patients with the definite symptoms, but the reliability coefficient needs to be enhanced.3. The study methods of proteomics on PBMC of SLE may make some contributions to pathogenesis study and foundation of new diagnosis criteria.