title: Gene expression changes in human islets exposed to type 1 diabetic serum.
abstract: A major obstacle to the success of islet cell transplantation as a standard treatment for labile type 1 diabetes mellitus is the immediate loss of up to 70% of the transplanted islet mass. Activation of the complement cascade and coagulation factors has been implicated in initiating the destruction of the islet graft. In this study, we analyzed the gene expression changes in islet cells following exposure to type 1 diabetes mellitus serum (T1DM). Isolated human pancreatic islet cells were cultured for 2 d to stabilize islet cell gene expression. Cultured islets were divided into three groups for treatment as follows: group 1 was treated with autologous donor serum, while groups two and three were treated with sera from ABO-matched allogeneic donors or autoantibody positive type 1 diabetic patient, respectively. Complement was detected using anti-C3 FITC and CH50 assay. Islet gene expression was analyzed using Illumina micro-array technology. Results were confirmed using real-time PCR. Immunofluorescent imaging demonstrated complement deposition only in the T1DM condition. Gene array and class prediction analysis generated a list of 50 genes that were able to predict the effect of T1DM serum on islets. Quantitative PCR corroborated microarray results. Both techniques demonstrated upregulation of MMP9 (243%), IL-1β (255%), IL-11 (220%), IL-12A (132%), RAD (343%) and a concomitant downregulation of IL-1RN (64%) in islets treated with T1DM serum. Islets treated with T1DM serum overexpressed genes associated with angiogenesis while decreasing transcription of genes that protect islets from inflammatory cytokines and reactive oxygen species.
authors.affiliation: Institute of Biomedical Studies, Baylor University, Waco, TX, USA.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Jackson
authors.forename: Mazhar A
authors.initials: MA
authors.lastname: Kanak
authors.forename: Ellen K
authors.initials: EK
authors.lastname: Grishman
authors.forename: Damien
authors.initials: D
authors.lastname: Chaussabel
authors.forename: Marlon F
authors.initials: MF
authors.lastname: Levy
authors.forename: Bashoo
authors.initials: B
authors.lastname: Naziruddin
mesh_terms: D001323:Autoantibodies / Q000097:blood; D003167:Complement Activation; D003176:Complement C3 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003922:Diabetes Mellitus, Type 1 / Q000097:blood; D020869:Gene Expression Profiling; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D006801:Humans; D017370:Interleukin-11 / Q000235:genetics; D053765:Interleukin-12 Subunit p35 / Q000235:genetics; D053583:Interleukin-1beta / Q000235:genetics; D007515:Islets of Langerhans / Q000276:immunology* / Q000378:metabolism*; D016381:Islets of Langerhans Transplantation; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics; D018631:ras Proteins / Q000235:genetics
chemical_list: D001323:Autoantibodies; D003176:Complement C3; C508858:IL12A protein, human; D017370:Interleukin-11; D053765:Interleukin-12 Subunit p35; D053583:Interleukin-1beta; C084355:RRAD protein, human; D020780:Matrix Metalloproteinase 9; D018631:ras Proteins