TSG-6 protein expression in the pancreatic islets of NOD mice.

The histologic hallmark of the development of type 1 diabetes (T1D) is insulitis, characterized by leukocytic infiltration of the pancreatic islets. The molecules controlling the early influx of leukocytes into the islets are poorly understood. Tumor necrosis factor alpha (TNFalpha)-stimulated gene 6 (TSG-6) is involved in inflammation, extracellular matrix formation, cell migration, and development. In the present study, we examined the expression and cellular localization of TSG-6 protein in islets of female non-obese diabetic (NOD) mice using frozen section immunofluorescence staining. Pancreata from nondiabetic (8 and 25 weeks old), prediabetic (230-280 mg/dl blood glucose) and diabetic (>300 mg/dl blood glucose) NOD mice were stained for TSG-6, insulin, CD3, CD11c, Mac3 and CD31. TSG-6 protein was detected in 67% of islets of prediabetic mice, 27% of islets of 25-week old nondiabetic mice, and less than 7% of islets of diabetic mice and 8-week old nondiabetic mice. Lastly, islet-derived TSG-6 protein was localized to the infiltrating CD3 and CD11c positive leukocytes.

The immunological synapse.

The immunological synapse plays a central role in organising the immune system. Through their synaptic activity both T and B cells usually, but not always, acquire the information that critically determines the level and nature of the responses that they make. For T cells much of that information comes from epicrine and paracrine cell-cell interactions in the cluster that forms around a dendritic cell. These interactions are being dissected by experiments in which two populations of TCR-transgenic T cells are combined in vivo. Another important aspect of synaptic activity is the way in which different levels of expression of MHC class II molecules influence Th1/Th2 balance. In exploring this form of control we are learning something of general importance about cis-regulation.

Early commitment of adoptively transferred CD4+ T cells following particle-mediated DNA vaccination: implications for the study of immunomodulation.

The early responses of CD4+ T cells to particle-mediated DNA immunisation were investigated using OVA-specific TCR-transgenic CD4+ T cells. Following adoptive transfer of these cells, mice were immunised by delivery into the skin of a plasmid encoding ovalbumin. Transgenic T cells underwent a rapid and transient antigen-specific activation, followed by clonal expansion (up to approximately 6% of total lymphocytes). Immunisation with ovalbumin in CFA evoked similar responses with slightly faster kinetics. Numerous antigen-specific T cells synthesising IFN-gamma (Th1) and IL-4 (Th2) were detectable using both intracellular staining and ELISPOT assays. This study provides a quantitative analysis of both T cell proliferation and Th1/Th2 balance following particle-mediated DNA immunisation and establishes a robust and sensitive model in which to assess modulation of helper T cell responses in DNA vaccination.