Figure 1

Blockade of TNF-α converting enzyme (TACE) activation decreased the anti-Ro/SSA autoantibodies (Abs)-induced expression of inflammation-related genes. To demonstrate the differential regulation of proinflammatory cytokines/chemokines and receptors gene expression in anti-Ro/SSA Abs plus TAPI-1-treated human salivary gland epithelial cells (SGECs), an inflammatory response and autoimmunity SuperArray was adopted. Panel a shows that anti-Ro/SSA Abs treatment determines the differential expression of genes in human SGECs in comparison with untreated control cells. The pooled total RNAs isolated from untreated and anti-Ro/SSA Abs-treated SGEC were used and real-time PCR arrays were run twice. From the expression of 84 key genes related to autoimmune and inflammatory response, 23 differentially expressed genes were identified as indicated. (Data represent the mean±s.e. of five independent experiments). Blockade of TACE activation with TAPI-1 reduces anti-Ro/SSA Abs-proinflammatory cytokines/chemokines induction. (b) Quantitative PCR validation of findings from superarray in panel a using primers and RNA samples from an independent experiment. The values represent the means of replicate samples and are normalized to housekeeping genes. (c) A human cytokine multi-analyte ELISA array was used to measure cytokine production by human SGEC treated with anti-Ro/SSA Abs plus TAPI-1. Values represent the optical density decrease versus anti-Ro/SSA Abs-treated SGEC cultures from triplicate wells. (Data represent the mean±s.e. of three independent experiments). (d) Intracellular distribution of interleukin (IL)-13, IL-17 and IL-22 in human SGECs treated with anti-Ro/SSA and/or TAPI-1 examined under fluorescence condition. A reduced immunoreactivity for cytokines expression was observed following TAPI-1 treatment (panels C, G, K). No fluorescence was detected in permeabilized cells treated only with secondary antibody-FITC (A, E, I).